Chronic Disease Priorities, Biomarker Breakthroughs, and CGT-RLT Synergies

Behind the Headlines, Episode 22

ICH Q12 and Post-Approval Changes

The quickly evolving development of biopharmaceuticals is impacting innovation in the industry overall.

Advancements in Biopharma Spur Innovation

What Pharma Manufacturers Need to Know

Every Pharmaceutical Manufacturing Professional Should Know

Patrick Lavery is an Editor for the MJH Life Sciences brands 

. Previously at MJH, he filled the same role for 

Orlando, Florida, USA - February 8, 2020: Hesperos headquarters in Orlando, Florida, USA. Hesperos is a biotechnology company that develops human-on-a-chip systems. | Image Credit: © JHVEPhoto - stock.adobe.com

 a biotechnology company with a focus on developing therapies for neurological disorders that are difficult to treat, and 

 an industry leader in organ-on-a-chip technology, have announced a strategic agreement for the purpose of accelerating preclinical development of Psilera’s lead compound that targets frontotemporal dementia (1).

Psilera and Hesperos have partnered to accelerate preclinical development of PSIL-006, a neuroplastogen candidate targeting frontotemporal dementia.

Hesperos’ organ-on-a-chip system uses patient-derived stem cells to model disease physiology, supporting data generation for FDA submissions.

FDA policy shifts promoting human-based lab models highlight organ-on-a-chip as a viable alternative to animal testing, aiding faster, safer drug development.

The compound, known as PSIL-006, integrates next-generation neuroplastogen, and the partnership with Hesperos will leverage that company’s advanced biological platform to develop a targeted treatment for unmet medical needs relating to frontotemporal dementia, a progressive neurological disorder for which treatment options are currently limited (1).

“Our collaboration with Hesperos represents a significant step forward in our mission to develop new treatments for neurodegenerative diseases,” said Chris Witowski, co-founder and CEO of Psilera, in a press release (1). “Utilizing their cutting-edge platform allows us to gain deeper insights into PSIL-006’s mechanism of action as we’re nearing first-in-human trials.”

The Hesperos platform, which integrates patient-derived induced pluripotent stem cells into interconnected, multi-organ systems, has been recognized for its ability to replicate human physiological responses, according to Hesperos, and has successfully supported multiple FDA Investigational New Drug and Orphan Drug Designation applications (1).

“We are excited to partner with Psilera as they advance next-generation neuroplastogens to potentially treat a range of hard-to-treat diseases,” said 

 PhD, co-founder and chief scientist at Hesperos, in the press release (1). “Our platform offers a unique opportunity to assess the compound’s effects on neural tissues derived from patients with neurodegeneration, potentially accelerating the development of a much-needed therapy utilizing clinically relevant functional readouts.”

Organ-on-a-chip technology was at the center of another partnership, 

 (2). Under that agreement, Pharmaron is tasked with validating CN Bio’s PhysioMimix technology across existing applications, while integrating organ-on-a-chip technologies into its R&D platform. Also in the deal, the partners agreed to explore potential new applications that would address unmet needs in drug discovery and development (3).

 as one of the human-based lab models that FDA would be shifting to, specifically those systems that mimic the heart and liver, in its objective to reduce and eventually replace animal testing in monoclonal antibody safety evaluations (4).

“For patients, it means a more efficient pipeline for novel treatments,” FDA Commissioner Marty Makary, PhD, said at the time (4). “It also means an added margin of safety, since human-based test systems may better predict real-world outcomes. For animal welfare, it represents a major step toward ending the use of laboratory animals in drug testing. Thousands of animals, including dogs and primates, could eventually be spared each year as these new methods take root.”

In the press release announcing the agreement between Hesperos and Psilera, that policy adjustment by FDA was cited as a means of promoting the benefits of organ-on-a-chip—for not only reducing the reliance on animal testing, but also enabling precise evaluation of drug efficacy and safety that has the potential to expedite clinic-ready datasets (1).

Psilera Collaborates with Drug Development Leader Hesperos to Advance Preclinical Modeling of PSIL-006 for Frontotemporal Dementia.

 Press Release. June 9, 2025.
2. CN Bio. 

CN Bio and Pharmaron Establish Long-Term Strategic Partnership to Develop OOC Technologies on a Global R&D Platform.

 Press Release. April 24, 2025.
3. Mirasol, F. 

CN Bio and Pharmaron Partner to Develop OOC Technologies for Global R&D.

FDA Announces Plan to Phase Out Animal Testing Requirement for Monoclonal Antibodies and Other Drugs.

Articles

The partnership leverages the Hesperos organ-on-a-chip platform in the preclinical development of Psilera’s lead compound targeting the progressive neurological disorder for which treatment options are few.

Hesperos, Psilera Agree to Accelerate Organ-on-a-Chip Treatment for Frontotemporal Dementia

Cynthia A. Challener is a contributing editor to Pharmaceutical Technology.

Efficient and effective delivery of complex and challenging drug substances is an ongoing issue facing developers of modern medicines. Delivery systems must overcome poor solubility and bioavailability while achieving targeted delivery to achieve maximum efficacy with lower doses while also minimizing unwanted side effects. Nanoscale technology is proving to be an attractive solution on all accounts, particularly for realizing targeted delivery into specific types of cells and across biological barriers.

Many of the unique properties of nanoscale delivery systems can be attributed to their small size (< 100 nanometers). At this scale, the surface area/volume ratio is high, leading to unique physical, chemical, and biologic properties. The ability to tune the surface properties of nanomaterials through functionalization further increases their utility for drug delivery, particularly with respect to their targeting capabilities. Nanoscale delivery systems can not only increase the bioavailability of poorly water-soluble drugs; they can also protect sensitive APIs from degradation and, when appropriately designed, remain stable in the bloodstream, allowing long-term sustained release and more consistent pharmacokinetics (1). The greater cellular uptake of nanoscale delivery systems also contributes to improved efficacy and reduced side effects.

Another important advantage of nanoscale delivery systems is the ability to deliver two or more APIs simultane

Nanoscale co-delivery systemsoffer improved performance for combination therapies because they deliver the different drug substances in a homogeneous manner (2). Ideal nanoscale carriers are stable, small, but able to support sufficient drug loading of both hydrophobic and hydrophilic APIs; protect the loaded drug from degradation; are suitably functionalized to exhibit a high binding affinity to target cells/ tissues; and have good biocompatibility.

Many different types of nanoscale delivery systems (e.g., lipidic, polymeric, inorganic, biomimetic) provide opportunities for designing solutions with high specificity for the particular API, disease target, and route of administration. In fact, they support the loading of most types of drug substances, including small molecules, proteins, nucleic acids, and diagnostic agents (1). They can also be customized in support of personalized medicines, and some have been developed that are capable of exhibiting both therapeutic and diagnostic activities (theranostics).

As more information is gained about the benefits of nanoscale delivery systems, researchers are exploring the applicability of a growing variety of nanoscale materials for this purpose (1–3). Nanoparticles have attracted particular attention, including those based on lipids, polymers (natural and synthetic), lipid-polymer hybrids, dendrimers, inorganic materials (metals, magnetic compounds, carbon nanotubes, silica-based systems, etc.). Other nanocarriers include nanocrystals; DNA-based systems; extracellular vesicles such as exosomes, microvesicles, apoptotic bodies, and phagosomes; nanoemulsions; and micelles.

Like any drug delivery technology, nanoscale systems are not perfect solutions and, depending on the particular nanotechnology, have different sets of limitations (1). Some of the nanomaterials used for drug delivery can, if the systems are not designed appropriately, pose toxicity and biocompatibility issues. One example is metallic nanoparticles if they accumulate in the body.

Similarly, while some nanoscale delivery systems can exhibit prolonged lifetimes in the bloodstream, others have short circulation times or can be viewed as foreign materials by the immune system and rapidly cleared. Some nanomaterials may also exhibit unexpected/unpredictable interactions with biological systems that lead to detrimental effects. These uncertainties and complexity of nanoscale drug delivery systems often lead to regulatory challenges.

The development of robust, predictable large-scale manufacturing processes can be a significant hurdle for some nanomaterials/nanoscale delivery systems. In addition, stability during storage can be an issue for some nanoscale systems, requiring careful formulation development. All these challenges tend to lead to high costs for therapeutics and vaccines leveraging nanoscale drug delivery systems.

Highly targeted delivery with nanoscale systems can be achieved using passive or active mechanisms. Passive delivery involves accumulation of the nanoscale system in the targeted cell/ tissue through enhanced permeability and retention (EPR), which is dictated by the shape, size, charge, and other surface properties of the unfunctionalized nanomaterial (4). This approach does not, however, completely reduce delivery to healthy cells, which can result in off-target effects.

Active targeting, meanwhile, is realized through functionalization of the surface of the nanocarrier system to both increase cellular uptake and ensure delivery to specific cells/tissues (4). Ligands are attached to the surfaces of the nanocarriers that have high binding affinity to receptors specific to the target cells. These ligands can include a range of compounds, including antibodies, aptamers, peptides, small molecules, and biomimetic materials such as cell membranes, among others. As an example, nanoscale delivery systems can be functionalized with PD-L1 monoclonal antibodies in anticancer therapies.

The targeting ability of nanoscale systems can benefit treatments for all disease classes, but this capability is of even greater importance in certain applications. Targeting of cancer cells is a major goal in oncology, as traditional systemic treatment result in severe side effects. The ability of nanocarriers to selectively attack targeted tumor cells allows for delivery of higher doses to improve efficacy while reducing cytotoxicity (1).

Therapeutic hyperthermia is a unique treatment made possible by use of nanoparticle delivery systems (1). Once the nanoparticles are localized in the tumor, they are exposed to laser radiation or an applied magnetic field, leading to an increase in temperature sufficient to kill the cancer cells. The improved targeting capability of nanoscale delivery systems also leads to improved activation of immune cells in cancer vaccines.

Nanoscale delivery systems are improving treatments for infectious diseases including human immunodeficiency virus (HIV) due to their targeting ability. Patient adherence with traditional medications is often low due to the high incidence of side effects. The high on-target bioavailability of nanoscale drugs thus has a measurable, positive impact.

Finally, nanoscale delivery systems are also improving the effectiveness of treatments for autoimmune diseases due to their improved targeting capability (1). Nanoparticles have been shown to deliver antigens and immunomodulators to antigen-presenting cells and lymphocytes with high specificity, and thus more efficiency, inducing antigen-specific tolerance 

Lipid nanoparticles (LNPs) have received significant attention in recent years as attractive systems for the delivery of nucleic acids for vaccine applications. Their success was clearly demonstrated with the approval of the two messenger RNA vaccines against COVID-19, according to McDavid Stilwell, CFO with CPTx. He notes, however, that some of the properties that make LNPs useful for vaccine applications (inflammatory behavior) also present substantial hurdles for systemic therapeutic delivery applications that require higher doses and for which adverse inflammation is undesirable.

There are also issues with targeting LNPs away from the liver, Stilwell observes. The field is working hard to overcome cell targeting challenges, but solutions are not expected soon. Stilwell notes that LNPs have now been successfully engineered to target immune cells, and CPTx is using this approach to deliver single-stranded DNA for its in vivo chimeric antigen receptor (CAR)-T development program.

In addition to ensuring targeted specificity and minimal off-target effects, key challenges with LNPs— and nanoscale delivery systems in general—include achieving consistent manufacturing scalability, controlling batch-to-batch variability, overcoming biological barriers (immune recognition, clearance, endosomal escape), and demonstrating robust safety profiles, adds CPTx’s CEO Hendrik Dietz. “Achieving reliable, reproducible, and cost-effective large-scale production remains critical, especially for transitioning from preclinical validation to clinical and commercial stages,” he says.

CPTx believes that DNA nanostructures presenting an immune-silent vector are a better alternative to currently available gene delivery technologies. “These nanocarriers are optimal for precision targeting, programmable drug release, and transient genetic expression, making them valuable for controlled gene editing and transient immunotherapies,” Stilwell contends. The goal, he adds, is to present a delivery solution that is not only technically superior, but that can also be manufactured in a cost-effective fashion.

The single-stranded DNA nanocarriers CPTx is developing are intended to enable direct, targeted in-vivo gene delivery. Scalable, high-purity DNA single- strand synthesis platforms, both for the gene encoding components of the nanoparticles and the helper material needed to build the nanocarriers, have been an important initial focus to ensure manufacturing challenges are not an issue, according to Dietz.

These new DNA nanostructures are precision engineered from therapeutic immune-silent DNA single-strands using a technique known as programmable DNA nanofabrication, uniquely combining precise control over composition and 3D shape, as well as delivery functionality. “The result is precise cell targeting without external carriers, bypassing issues associated with existing delivery technologies such as viral vectors and lipid nanoparticles, including manufacturing hurdles, immunogenicity (in the case of viral vectors), and targeting challenges,” Dietz states.

The DNA nanocarriers “integrate therapeutic payloads and delivery functionality into rationally engineered particles designed to realize episomal, non-integrated gene expression in target cells or tissues with minimal side effects,” Dietz explains. Functionalization in a site-specific fashion with molecular targeting motifs promotes highly selective binding to specific cells or tissues and efficient cellular uptake, he says.

The modular approach also allows for simultaneous delivery of multiple therapeutic gene payloads and/or simultaneous targeting of multiple tissues or cell types, enhancing effectiveness, safety, and control compared to traditional approaches, according to Dietz.

CPTx is currently working to improve the delivery efficiency in various prototypes while also validating safety and efficacy in animal models with the goal of licensing this DNA nanocarrier delivery solution to drug developers with established payload genes and target tissues.

1. Tenchov, R. et al. Transforming Medicine: Cutting-Edge Applications of Nanoscale Materials in Drug Delivery. 

. 2025 19 (4). https://pubs.acs.org/ doi/10.1021/acsnano.4c09566

2. Sun, L. et al. Better Together: Nanoscale Co-Delivery Systems of Therapeutic Agents for High-Performance Cancer Therapy. F

ront. Pharmacol., Sec. Pharmacol. Anti-Cancer Drugs

. 2024 Vol. 15. DOI: 10.3389/fphar.2024.1389922.

3. Li, Y.; et al. A Nanoscale Natural Drug Delivery System For Targeted Drug Delivery Against Ovarian Cancer: Action Mechanism, Application Enlightenment And Future Potential. 

Front. Immunol., Sec. Canc. Immun. & Immunother.

, 2024 Vol. 15. https://doi.org/10.3389/\ fimmu.2024.1427573

4. Fan, L.; Wang, A.; and Shi, D. Targeted Nanoscale Drug Delivery Systems For Melanoma Therapy. 

. 2023 Vol. 86, 104724. https://doi. org/10.1016/j.jddst.2023.104724

Unique surface properties and ligand functionalization provide real advantages.

Achieving Targeted Delivery with Nanoscale Systems

cold storage room with shelves stocked with boxes and containers. Cooling fans and a digital temperature control panel ensure precise climate conditions. refrigeration - storage shelves | Image Credit: © Grispb - stock.adobe.com

The global contract packaging organization 

 is announcing an expansion of its contract packaging and cold chain storage services in both the United States and The Netherlands (1). The company intends to scale its logistics infrastructure to meet rising demand it has seen for injectables and biologics, as well as bolster its supply chain resilience and deliver faster turnaround times and greater flexibility for clients.

In a press release on June 5, 2025, Tjoapack detailed the changes planned for the facilities in Clinton, Tenn., in the US, and at Etten-Leur in The Netherlands.

Tjoapack is expanding packaging and cold chain services in the US and EU to support growing demand for biologics, injectables, and self-administered therapies.

New automated lines in Clinton, Tenn., will enable high-throughput vial and auto-injector packaging with integrated traceability, labeling, and serialization.

Etten-Leur now offers fully automated prefilled syringe packaging, as both sites scale cold storage to maintain GMP standards across temperature-sensitive products.

At the Clinton site, Tjoapack is completing installation of two advanced packaging lines that will support high-throughput, sterile packaging operations (1). A line expected to be completed in the second quarter of 2025 is high-speed and fully automated for vial packaging, with an annual processing capability of 20 million vials—including integrated labeling, cartooning, and serialization. In the third quarter, a semi-automated auto-injector line that offers end-to-end traceability and labeling will be scheduled for qualification. Cold storage capacity at this site has been expanded and validated, providing a total of 160 good manufacturing practice-compliant pallet spaces in controlled 2–8°C conditions.

The Etten-Leur facility has also seen its cold chain capacity increase, bringing on 84 additional pallet spaces to push its total to 120, under the same temperature guidelines. This site, according to Tjoapack, additionally has completed its first commercial project for prefilled syringe (PFS) packaging (1). The fully automated PFS line offers syringe assembly with or without safety devices, automated plunger rod insertion, and insertion of ancillary components like needles, among other features.

“These upgrades directly address the accelerating needs of our clients in the biologics and sterile injectable space,” said 

 CEO of Tjoapack, in the press release (1). “We are committed to building scalable, compliant infrastructure that ensures the efficient delivery of vital therapies without compromising on flexibility or quality. By strengthening its injectable packaging and cold chain capabilities, Tjoapack is supporting a wider range of primary container formats, including vials, ampoules, syringes, and auto-injectors, while scaling capacity to better serve both new and existing clients across North America and Europe.”

®, Joe Compton, senior director of Business Development and Alliance Management for Tjoapack US, said 

 would be driven in part by the rise of biologics self-administration.

“The development of PFS and auto-injectors has simplified the injection process, making it easier for patients to self-administer biologics at home,” Compton said (2). “Auto-injectors are designed for intuitive use, often requiring a single step to deliver the medication. This eliminates the complexity of traditional injections, which involve multiple steps like preparing the syringe, measuring the dose, and administering the injections. Auto-injectors feature ergonomic designs and clear instructions, making them easy to handle and operate, even for patients with limited dexterity or those unfamiliar with injections.”

At the Drug, Chemical & Associated Technologies Association 

® Group about recent innovations in biopharmaceutical manufacturing, and Tjoapack’s strategy in responding to those trends.

“We've been investing heavily in expanding our cold chain capabilities, in expanding our capability and capacity to package unique configuration for injectable products, whether that is a product that comes to us in a syringe format, a vial format, or an auto injector format,” Belden said (3). “[We have] the ability to either automate or semi-automate, label, and then finish [to] pack those products while maintaining tight control of time [and] temperature to allow for packaging, and then getting those materials and finished goods back into cold storage.”

The full interview with Belden can be seen 

1. Tjoapack. Tjoapack Continues to Expand Its Global Infrastructure with Strategic Investments in Injectable Packaging and Cold Chain Capabilities. Press Release. June 5, 2025.
2. Compton, J. 

Meeting Secondary Packaging Challenges of Innovative Delivery Systems for Biologics.

Most of the company’s new additions, if not already online, will be scheduled for commissioning, validation, or qualification before the fourth quarter of 2025.

Tjoapack Announces Packaging, Cold Chain Expansions in US and Netherlands

Blue and white capsules pill in blister pack arranged with beautiful pattern. Global healthcare concept. Antibiotics drug resistance. Antimicrobial capsule pills. Pharmaceutical industry. | Image Credit: © Artinun - stock.adobe.com

At conferences and trade shows up and down the calendar, bio/pharmaceutical companies take the opportunity to showcase new, updated, or redesigned equipment, responding to not only the latest demands of the market, but also the innovations of competitors. In this interview with 

®, Sheikh Akbar Ali, PhD, general manager and head of Development and Technology for ACG Packaging Materials, reviews the state of equipment innovation, the emerging technologies that are shaping a new standard of efficiency, how global pressures are affecting the supply chain for raw materials, and what options are available to manufacturers who desire to make their processes more sustainable.

 To start, what are some general trends you are seeing in equipment? Smart tools and three-dimensional (3D) printing are two specific areas in which we have observed progress so far in 2025, and artificial intelligence (AI) continues to be a growing factor in all types of processes. What is most in demand at the current time?

 One growing trend in pharmaceutical packaging is the move toward digital and laser printing technologies. Digital, or on-demand, printing is gaining popularity for enabling batch-level information to be printed just before medicines are packed. This supports traceability, eliminates errors, and increases patient safety as well as flexibility, especially in smaller or personalized medicine batches. Laser printing is also emerging, offering a high-energy, ink free alternative for durable and tamper-evident markings. ACG has been investing significantly in AI to develop smarter equipment. As an example, our vision inspection systems are capable of detecting even the slightest manufacturing defects, ensuring only flawless products are produced. The integration of AI has resulted in a quantum leap forward in defect-detection accuracy, particularly for low-contrast product and foil combinations. Also, training these systems now requires minimal human involvement, with the machine continuously learning and refining its performance through deep learning, and improving its own efficiencies incrementally.

 Many companies are taking steps to increase capacity to meet the demands of their customers. Is there a category of equipment that companies are tending toward including in their expansion plans?

 Like many other sectors, pharmaceutical companies are highly attuned to cost efficiency and inventory management. With a growing emphasis on sustainability, they are now equally attentive to the carbon footprint of their products. As a result, packaging companies are focusing not just on capacity building, but on developing capabilities to produce packaging with the lowest possible environmental impact. Particular attention is being given to solvent-free adhesives and sealing lacquers. Coating and laminating lines that accommodate these solvent-free materials are becoming increasingly valuable. Energy efficiency is another major consideration. Technologies that capture waste heat from the process stack and reuse it to reduce overall energy consumption are gaining traction. And the demand for sustainable packaging materials has accelerated the need for advanced blistering lines, and systems that are faster, modular, and reconfigurable with minimal change over time. These innovations enable manufacturers to efficiently handle a variety of eco-friendly materials without compromising on productivity or quality. In addition, companies setting up new production lines or facilities are prioritizing digital and on-demand printing infrastructure. This enables greater operational flexibility and supports compliance with evolving regulatory requirements.

 How have outsourcing relationships and partnerships evolved to meet the manufacturing needs of all concerned—especially given the currently tenuous geopolitical climate?

 Global supply chain disruptions—driven by geopolitical tensions—are significantly affecting raw material sourcing and timely delivery of packaging materials to pharma companies. This creates bottlenecks in both production and delivery. The issue is particularly severe for sustainable materials needed to meet net-zero goals, as these materials are either scarce or unaffordable.

 What about single-use technology? Where do you foresee the next stage of innovation in that area, and how does that align with sustainability goals for the industry?

 In pharmaceutical applications, reusing packaging is not feasible due to contamination risks. Therefore, single use remains the industry standard. However, significant innovation is occurring to align single-use packaging with sustainability goals. Key developments include, firstly, polyvinyl chloride (PVC)- and halogen-free alternatives in thermoforming and cold forming applications, such as thermoform blisters where the PVC is replaced with APET (amorphous polyethylene terephthalate), PP (polypropylene), or PE (polyethylene); cold form blisters where the PVC is replaced with PP, PE, or BOPET (biaxially oriented polyethylene terephthalate), lidding foil where the halogenated primers and heat seal lacquers (HSLs) are replaced with halogen-free alternatives, or universal sealing lacquer instead of specific lacquer for specific sealing layer. A second key development is in the reduction of mass, evidenced by a compact blister where the blister dimension has been reduced to accommodate the increased number of blisters per stroke of the blister line. A third development is in recyclable materials, namely thermoforming material where both the forming and lidding material are made of the same material and can be recycled in existing recycling infrastructure. And the last is in biodegradable and compostable materials—biodegradable PVC, which is certified to degrade faster than the regular PVC, and paper-based blister packs, which are made of certified compostable materials. These developments enhance recyclability, minimize material usage, and offer safer disposal options, bringing sustainability to single-use packaging without compromising either safety or functionality.

 Are there any new, imminent, or expected regulatory changes that companies and their clients should be aware of?

 There is a notable gap in global regulations for alternative sustainable packaging materials. Current standards primarily cover traditional materials like PVC, which hampers the adoption of newer, eco-friendly options. This regulatory vacuum makes it difficult for pharmaceutical companies to switch to sustainable formats, despite technological readiness. Additionally, the European Union (EU)’s Carbon Border Adjustment Mechanism is a pressing concern. It imposes a tax on imported products, increasing the cost of non-EU packaging materials. This may impact pricing and competitiveness for companies like ACG that export into the EU market.

®
Vol. 49, No. 5
June 2025
Pages: 20–21

When referring to this article, please cite it as Lavery, P. How Smart Technology is Helping Reach Sustainability Goals in Drug Packaging. 

Smart technologies, such as digital and laser printing, incorporated into manufacturing equipment are helping manufacturers align with sustainability goals, according to Sheikh Akbar Ali, general manager and head of Development and Technology for ACG Packaging Materials.

How Smart Technology Is Helping Reach Sustainability Goals in Drug Packaging

Automatic machine for filling ampoules and vials. Modern pharmaceutical equipment. | Image Credit: © nordroden - stock.adobe.com

TurboFil Packaging Solutions is introducing its newest piece of equipment, called the MediFil Cart (1). In a press release on June 4, 2025, the company described this as a standalone, fully servo-controlled ceramic vial filler system, designed as a cart-mounted module that provides convenience and ease of portability between lines.

MediFil Cart is operator-friendly, flexible, and precise

TurboFil’s new MediFil Cart offers precise, servo-controlled vial filling with up to 12 individually optimized channels and easy portability between lines.

The system’s ergonomic, cart-mounted design supports remote control, rapid setup, and integration into existing lines to boost uptime and operational efficiency.

TurboFil emphasizes balanced automation—while automated filling enhances accuracy, sterility, and inspection challenges must also be considered at scale.

The compact system has the potential to come equipped with up to 12 piston pumps, with each of the filling channels driven by its own motor drive, according to TurboFil (1). This design allows for each channel to be individually optimized for best accuracy, while a suck-back feature is designed to minimize drips. Stainless steel pump housing is intended for ease of disassembly and reassembly. Overall, the company said, the system offers an ergonomic design that is operator-friendly, flexibility for production floors, and tight-tolerance dosing precision.

Additionally, TurboFil said the system is easy to set up and affords control over all fill parameters—enable/disable, volume, speed, acceleration, and suck-back (1). Ethernet connectivity can provide either remote system control or integration into larger production lines.

“As both filling tolerances and production scheduling windows grow ever tighter, pharma manufacturers are seeking solutions that combine precision with expedience and flexibility,” said 

, vice-president of Marketing & Business Development for TurboFil Packaging Machines, said in a company press release (1). “The MediFil Cart is designed as a versatile filler that allows operators to perform set up removed from a production line, then integrate the unit in a just-in-time fashion that maximizes uptime and overall efficiency.”

Fully automated vs semi-automated operations

® Group got some perspective from TurboFil on automating operations, particularly the distinction between fully automated and semi-automated approaches, 

 of packaging trends. At the time, the company was promoting a custom-built benchtop draw filler, which offered better accuracy and a less labor-intensive setup than manual filling (2).

Even while touting the advantages of that system, however, Smook cautioned against an overreliance on automated practices.

“Automation is rarely as simple as devising a machine to replace a manual process,” she said in the article, which was published in both 

®. “Adjacent issues such as sterility and inspection, which can become more challenging at higher line speeds, also must be addressed.”

TurboFil is now adding the MediFil cart to its broad portfolio of standardized equipment, and the company says it also frequently collaborates with customers in the design, development, and construction of customized, built-to-spec machines for the pharmaceutical, medical device, health & beauty, and chemical industries (1).

For more on equipment trends, keep an eye out for the June 2025 edition of 

, in which Sheikh Akbar Ali, general manager and head of Development and Technology for ACG Packaging Materials, reviews the state of equipment innovation, the emerging technologies that are shaping a new standard of efficiency, how global pressures are affecting the supply chain for raw materials, and what options are available to manufacturers who desire to make their processes more sustainable (3).

1. TurboFil Packaging Machines. TurboFil Introduces Versatile Multiheaded Vial Filler Cart with Servo-Controlled Ceramic Pumps. Press Release. June 4, 2025.
2. Forcinio, H. 

How Smart Technology Is Helping Reach Sustainability Goals in Drug Packaging.

The company describes the compact system as ergonomically designed, providing precision dosing and production floor flexibility.

TurboFil Introduces MediFil Cart as Standalone, Servo-Controlled Ceramic Vial Filler

Richard Easton, PhD, is Technical Director–Structural Analysis, BioPharmaSpec Ltd,

Christopher Ziegenfuss is Vice President, at BioPharmaSpec, Inc

A magnifying glass examining a vintage vaccine vial surrounded by four minimalistic chart icons, symbolizing healthcare, medicine, and analytics in the context of global health monitoring. | Image source: ©huiying - stock.adobe.com

Biopharmaceutical product development is burgeoning with developments in treatment approaches, chemistries, and vectors giving companies new strategies for targeting disease. Biosimilars continue to build as a sector of this market with an estimated 2025 value in the region of US$42 billion (1). Many other products are also under development with innovative designs in the areas of fusion proteins, antibody drug conjugates (ADCs) and related species, peptides, and oligonucleotides.

No matter the nature of the product under development, one of the single most important factors during that development phase is the analytical characterization of the product. The need to characterize goes beyond the obvious requirement for demonstration of correct structure of the API itself but also demonstrates control of the manufacturing process, assesses impurity profiles, and ultimately feeds back to changes and improvements in the manufacturing process to ensure the best quality product. Characterization data also builds in quality to the product by having that deep knowledge of its structure and compositional profile. A detailed and thorough characterization is required to be presented to regulatory authorities as part of any submission for regulatory approval and marketing authorization.

The expectations for structural analysis are clearly laid out in a guideline published a number of years ago by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). ICH Q6B provides a clear framework for expectations regarding the structural characterization of biopharmaceutical products (2). While the document itself is a quarter of a century old at this point, it is laid out in such a way that relevancy is maintained. It uses terms like “to the extent possible”, thus allowing for methodological developments pushing the ability to detect structural details in ever greater and more precise ways. That being said, it is recognized that an update is due to acknowledge and bring into focus current analytical practices and to align it with other ICH guidelines. This guideline forms the key document referenced by regulatory authority documents that detail structural characterization requirements of biopharmaceuticals. This covers new biotechnologically derived protein, glycoprotein, and peptide products as well as biosimilars (3, 4). So why is ICH Q6B so important, and what does it outline?

Biopharmaceuticals, which are produced either in cellular systems or created through chemical synthesis, are molecules with incredible complexity. They can be orders of magnitude larger than what we may consider basic pharmaceuticals to be and with this increased size comes not only increased complexity but increased ways in which this complexity manifests. One must consider not only the basic linear structure of the molecule but also the three-dimensional (3D) shape, modifications (either wanted or unwanted) that occur on the molecule, and interactions that may occur within and between molecules themselves (e.g., disulfide bridges and aggregation). Also, consideration must be given to impurities that may arise as a result of the manufacturing or purification processes. All these must be investigated to give as a full a structural characterization of the product as possible and also to use the data as a tool to hone the manufacturing process itself to produce the best quality drug possible. ICH Q6B details the aspects of molecular structure for which characterization is expected.

Firstly, and perhaps most importantly, ICH Q6B requires that the primary amino acid sequence of the product (protein or polypeptide) is determined. This is crucial because a recombinant protein or synthesized peptide must have a known and specific amino acid sequence to fulfill its function. No precise methodology is given for how this should be performed, but modern methods of sequencing rely heavily on state-of-the-art mass spectrometry to provide the most complete and accurate data. Depending on the instrument type, mass spectrometers (e.g., quadrupole time-of-flight [Q-TOF] geometry and similar instruments) can generate mass values with very high mass accuracy (low ppm). This high mass accuracy in combination with this type of instruments’ ability to fragment peptides generates precise sequence information.

Proteins are proteolytically digested to peptides using strategies tailored to the protein under investigation. Peptides are separated and analyzed by on-line liquid chromatography–mass spectrometry (LC-MS), and fragmentation data generated are then used to determine peptide and ultimately protein sequence. This is not to say that MS will be able to generate 100% of the sequence coverage. There may be regions of the protein where peptide generation is difficult or fragment ions are weak or ambiguous. In these instances, gas phase sequencing of purified peptides (also known as Edman sequencing) can be performed to sequentially identify amino acids based on their chromatographic elution positions following specific, sequential chemical labeling.

ICH Q6B also requires that the overall amino acid composition of the product is investigated. To achieve this, the relative molar amounts of each amino acid in the product need to be determined. This is typically carried out using relative quantitation of amino acids following acid hydrolysis of the product and chromatographic separation of the released and derivatized amino acids. Comparison to known amounts of standard amino acids allows for accurate quantitation of each amino acid. It is recognized that no single acid hydrolysis condition will be suitable for all amino acids, so this must be taken into account in terms of recoveries of amino acids and if necessary alternate hydrolytic approaches taken.

Both the N- and C-termini of the product are required to be assessed to demonstrate the degree of homogeneity in each case. Post-translational processing may result in changes to the termini (e.g., ragged ends or N-terminal pyroglutamination), and these must be investigated. N-terminal analysis can be performed by gas phase sequencing if there is a free amine group at the N-terminus (e.g., no pyroglutamination or other blocking group), but there is no equivalent analysis for the C-terminus. Therefore, these terminal investigations are often performed as part of the peptide mapping study.

Peptide mapping is one of the most data-rich analyses that ICH Q6B requires. This investigation is, in some ways, associated with the procedure for primary amino acid sequencing discussed previously. However, peptide mapping focuses on mass analysis of proteolytically released peptides and comparison to the expected masses. The use of high-end mass spectrometers that can generate real-time fragmentation data allows a further confirmation of the identity of peptides detected. Peptide mapping generates a wealth of data, because the combination of specific proteolytic digest and on-line LC-MS analysis of the digest products allows not only accurate mass identification of the peptides themselves but also the identification of any post translational modifications (

). These may either be specific to the production process itself (such as pyroglutamination of the N-terminus as is frequently seen in monoclonal antibodies (mAbs) or may be the result of purification processes (such as amino acid oxidation or deamidation events) and may indicate that process change is required. Peptide mapping also generates data that are required for other areas of ICH Q6B, specifically terminal amino acid sequence identification and disulfide bridge analysis. It also identifies glycosylation sites and can be used to assess the degree of glycosylation site occupancy for glycoproteins. It thus fits well into a multi-attribute monitoring workflow.

Figure 1: Examples of the most commonly found post-translational

modifications on proteins. These can be identified and investigated through

peptide mapping studies with detailed glycan characterization taking place

through specific studies. (Figure courtesy of the authors)

Disulfide bridges are links formed across a protein chain, or between protein chains, via the side chain thiol groups of the amino acid cysteine. These bridges serve to maintain the correct 3D shape of the molecule and are very precisely formed, such that the same bridges will be formed from the same pair of cysteine residues in each copy of the protein. The disulfide bridges in a protein are required to be identified to confirm that first they are present and secondly that they are correctly formed. The presence of incorrect disulfide bridges is termed “scrambling” and indicates the presence of a misfolded protein population. This population is likely to result in inactive material but could at worst lead to an immunogenic response in the patient or product aggregation. Disulfide bridges can be identified through targeted proteolytic and/or chemical digestion strategies and MS-based analyses and can be seen as akin to peptide mapping, albeit focused specifically on the bridges (

). It is also important to assess for any unbridged cysteine residues.

Figure 2: Disulfide bridge analysis. Bridged peptides are identified by mass following proteolytic digestion. Fragment ions or a reduction step are then used to confirm the bridge identification. (Figure courtesy of the authors)

Many of the biopharmaceutical products on the market today are glycoproteins with mAbs being the most widespread example. For glycosylated products, it is a requirement that the sugar structures of the molecule (known collectively as glycosylation) are thoroughly characterized. This requires a quantitative analysis of the individual monosaccharides in the glycans (which can be performed by gas chromatography [GC]-MS or LC with fluorescence detection for sialic acids), but most significantly the overall glycan profile must be investigated. This usually involves release of the glycans, and either fluorescent labeling and analysis by on-line LC-MS with fluorescence detection (

) or MS analysis of released glycans following chemical derivatization, depending on the type of glycan being analyzed. This process allows the relative abundances of each glycan to be assessed along with the identification of each glycan’s composition through mass values detected.

Figure 3: Fluorescence profile of released N-glycans from a monoclonal antibody following fluorescent labelling and liquid chromatography-mass spectrometry (LC-MS) analysis. MS provides mass information allowing identification of N-glycan compositions. (Figure courtesy of the authors)

To be able to fully define their structures, the linkages between the monosaccharides must also be characterized. Linkage analysis can be performed through a series of elegant chemical steps culminating in GC-MS analysis of the end products. The elution positions and fragmentation patterns identify the original linkage and type of each monosaccharide. Knowing the biosynthetic pathways of glycosylation along with the data generated, it is then possible to define the population of glycans on the product to the extent possible. ICH Q6B also requires that, for glycoproteins with more than one glycosylation site, the glycan profile at each site must also be characterized. Glycosylation is not a templated post-translational modification (PTM) and can be highly heterogenous, depending on the glycoprotein. A detailed knowledge of glycan biosynthesis is needed to fully interpret the data and define the glycan structures indicated by the data.

Modern MS instruments have very high mass accuracy and resolution, allowing an accurate assessment of the molecular weight of the intact product and associated heterogeneity (low ppm). It not only provides data related to the size of the molecule but also may provide orthogonal data supporting other primary assessments of key molecular characteristics including post-translational modifications. Analysis under various conditions (e.g., native [

], reduced, and/or following deglycosylation) allows for an assessment of the overall product mass as well as individual protein chains in multimeric species and protein chains in the absence of glycan heterogeneity.

Figure 4: Intact molecular weight of a sample of Herceptin (trastuzumab). On-line liquid chromatography-mass spectrometry (LC-MS) of the sample was performed followed by transformation of the raw mass data to generate the intact mass values. The mass differences observed between the major signals are consistent with differing levels of galactose on the N-glycans. Minor signals indicate that other sources of heterogeneity exist in the sample,and these can be further investigated. (Figure courtesy of the authors)

The MS data should be used alongside data from more traditional methods such as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (or its more modern capillary-based equivalent of capillary electrophoresis-SDS) and size-exclusion chromatography (SEC) when compiling a robust characterization package, to generate orthogonal data sets. With their technical and methodological improvements, advanced techniques provide valuable information on overall product mass profiles. With properly designed experiments, these analyses provide data that will be indicative of the quality of the manufacturing process and the product itself.

Charge isoform analysis provides a profile of species within the sample based on separation of the various charge states. There are many structural attributes within a protein that can affect the charge profile, the most frequently encountered being the post-translational modification deamidation, the presence of sialic acid in glycoproteins, or in the case of monoclonal antibodies (mAbs) the presence of pyroglutamate or heavy chain C-terminal lysine.

ICH Q6B notes the isoform pattern can be assessed using isoelectric focusing or other appropriate techniques. Frequently the capillary-based technique of image capillary isoelectric focusing (icIEF) is used due to its speed, resolution, reproducibility, and reliability of UV and fluorescence-based detection for relative quantitation. Data generated gives charge profile information, which can be further investigated using specific sample treatments prior to reanalysis. This is another area where collection of orthogonal data builds a thorough understanding of the product. For example, charge-based PTMs detected during peptide mapping or glycan analysis experiments may give rise to unique isoform profiles. Along with isoelectric focusing, ion-exchange chromatography can be used to assess isoforms as well as chromatographic patterns.

It is vital to be sure of the amount of product being administered to ensure patient safety and for product bioassay data generation. A common UV absorption reading can be used to measure protein content. When this strategy is applied, it is critical to determine the extinction coefficient. This determination can be achieved using a combination of ultraviolet-visible spectroscopy spectrophotometry and optimized amino acid analysis that has been assessed specifically on the product to demonstrate its suitability. Very often this encompasses a qualification or validation of the method to ICH Q2(R2) specifications (5,6).

Some aspects of structural characterization requirements specified by ICH Q6B overlap with one another. In the case of electrophoretic patterns for example, the same icIEF and CE-SDS experiments applied under the headings of size and/or isoform patterns can be used to provide the required data on identity, homogeneity, and purity for the characterization dossier that this analytical section covers.

The identity, homogeneity, and purity of biologic products needs to be evaluated, and this can be carried out using a variety of chromatographic techniques. The most commonly employed are reverse-phase, SEC, and ion exchange chromatography, each evaluating a different physicochemical characteristic of the molecule. As with other characterization assays, additional or substitute methods such as HIC or HILIC (hydrophilic interaction chromatography), can be used to enhance knowledge and understanding of the product depending on its structural characteristics. Several chromatographic techniques are expected to be applied to separate out components based on different physical properties such as charge, hydrophobicity, and mass. This gives the most comprehensive investigation into the overall composition of the sample because any coeluting species in one technique will likely separate in others.

Spectroscopic techniques are applied during characterization to assess the higher order (secondary and tertiary) structure. The 3D shape of a biological product plays a critical role in its function and therefore must be characterized to the extent possible. The higher order structures of molecules can be composed of different types of structural features, depending on the nature of the primary amino acid sequence and how it interacts with itself in its solution environment to produce the final 3D shape. At the secondary structural level, ordered structural features such as alpha helices or beta sheet structures can be produced, depending on the amino acid sequence, but regions of more random-type structure can also be found. How these secondary structural units interact with one another and assemble in three dimensions gives rise to the tertiary level of molecular structure. Various techniques should be applied during characterization, each of which has its own relative strengths and will thus probe the structure in different and orthogonal ways. The techniques of circular dichroism (CD), Fourier transform infrared spectroscopy (FT-IR), fluorescence analysis, microfluidic modulation spectroscopy (MMS) and nuclear magnetic resonance (NMR, both 1D and 2D) are all appropriate for higher order structural (HOS) analysis and provide a meaningful orthogonal dataset (7).

An assessment of aggregation is important as part of an investigation into product related impurities, which is also covered by ICH Q6B. Orthogonality is again important here and the application of two techniques such as SEC-multi-angle light scattering (MALS) (a column-based method, 

) and SV-AUC (a non-column-based method), which work on different physical principles, gives meaningful data.

Figure 5: Size exclusion chromatography-multi-angle light scattering (SEC-MALS) analysis of a protein showing the presence of mostly monomer (peak eluting at approx. 6 mins) but other higher mass species are also detected such as dimer, and trimer eluting at approx. 5.5 mins and 5.0 mins respectively. Lower levels of other higher oligomeric states are also present. The UV profile is shown with the MALS mass data given a discreet lines across peaks in the upper portion of the image. (Figure courtesy of the authors)

The ICH Q6B guidance document has been adopted by regulatory agencies due to its comprehensive coverage of structural requirements both for the product itself but also for impurities found within it. These in-depth characterization studies must be performed during product development and following significant process changes and are expected to be present in any drug application document.

The nature of biologically produced biopharmaceuticals inevitably results in some level of heterogeneity that must be assessed and for which techniques must be used that can provide clear and detailed analytical data. Researchers who follow the suggested guidelines outlined in ICH Q6B, using state-of-the-art analytical equipment and techniques, will be well-positioned to move their product through the regulatory processes and also for critically assessing the production process itself.

Mordor Intelligence. Biosimilars Market Size | Industry Growth & Forecast Report. Mordorintelligence.com (accessed May 5, 2025).

ICH. Specifications. Q6B. Test Procedures and Acceptance Criteria for Biotechnological/Biological Products (ICH, March 1999).

FDA. Development of Therapeutic Protein Biosimilars: Comparative Analytical Assessment and Other Quality-Related Considerations, Draft Guidance for Industry (CDER, CBER, 2019).

EMA. Guideline on Similar Biological Medicinal Products Containing Biotechnology-derived Proteins as Active Substances: Quality Issues (revision 1) (May 22, 2014).

ICH. Q2(R2). Analytical Validation (ICH, 2023).

Easton, R.L. Structural Characterization Methods for Biosimilars: Fit-for-Purpose, Qualified, or Validated. GaBI J., 2022 11, 41-44

Greer, F and Easton, R.L. Biosimilars–Increasing Regulatory Focus on Orthogonal Analytical Characterization. International Biopharmaceutical Industry, 2021 4, 10-13.

Richard Easton, PhD, is Technical Director–Structural Analysis, BioPharmaSpec Ltd, and Christopher Ziegenfuss is Vice President, at BioPharmaSpec, Inc.

®
Volume 49, No. 5
June 2025
Page: 22–28

When referring to this article, please cite it as Easton, R. and Ziengenfuss, R. ICH Q6B for Analytics. Pharmaceutical Technology 2025 49 (5).

ICH Q6B provides expectations and a clear framework for the structural characterization of biopharmaceutical products. 

ICH Q6B for Analytics

Adam Covitt discusses Federal Equipment Company's innovative initiatives, including PharmParts and Techceuticals

Training Centers and eCommerce: A Look Into Federal Equipment Company's Pharma Innovations

Adam Covitt, President of Federal Equipment Company, shares exciting updates from Techceuticals—the company’s state-of-the-art training center in Cleveland, Ohio—where new technologies are enhancing hands-on learning and client engagement. He also discusses PharmParts, a streamlined eCommerce platform designed to help CDMOs quickly source critical spare parts with next-day delivery. Learn more about:

Techceuticals and PharmParts, value-add businesses that focus on novel technologies, comprehensive training, and simplicity for CDMOs.

How the trade tariffs are fueling the shift back to oral solid dosage (OSD) products like tablets and capsules.

How online industrial auctions are giving pharma manufacturers a smarter way to monetize surplus equipment and reinvest in future-ready technologies.

Videos

How Techceuticals, PharmParts, and smart auction strategies are ushering in a new era of efficiency and innovation for CDMOs 

Training Centers and eCommerce: A Look Into Federal Equipment Company’s Pharma Innovations

Three groups of people are connected by lines. Coordination and knowledge sharing. Outsourcing. | Image Credit: © Uladzislau - stock.adobe.com

The role of contract development and manufacturing organizations (CDMOs) is at an inflection point at the midpoint of the 2020s, mainly taking into account the impacts of two seismic factors: geopolitical shifts, driven in large part by the tariffs enacted in the first several months of United States President Donald Trump’s second term, and the seemingly omnipresent prevalence of artificial intelligence (AI).

The versatility provided by CDMOs is, by necessity, changing with the times. At the Drug, Chemical & Associated Technologies Association (DCAT) Week 2025, India-based Shilpa Medicare said it was launching a newly formed, full-service, “hybrid” CDMO aiming to serve both small- and large-molecule customers in addition to peptides, with a therapeutic specialization in oncology (1,2). Shilpa said the hybrid model would adopt a dual approach: offering comprehensive discovery, clinical, and commercial outsourcing services, but also including commercially ready, “off-the-shelf” novel formulations for business-to-business licensing.

In April 2025, a new CDMO, Meribel Pharma Solutions, was announced to be headquartered in the United Kingdom (3,4). The creation of this mid-sized CDMO was made possible by an acquisition of Synerlab Group, another CDMO, combined with the securing of seven manufacturing sites in Europe formerly belonging to Recipharm. The new company offers 10 manufacturing sites and three drug development services sites in France, Spain, and Sweden, and has established centers of excellence in drug development, lyophilization, preservative-free multidose technologies, and flexible stick-pack and sachet production, meant to support both human and veterinary health products while operating under stringent global regulatory certifications.

One initiative that had been announced prior to the start of 2025 was Thermo Fisher Scientific’s Accelerator Drug Development service, described at the time of its October 2024 launch as the company’s 360° CDMO and contract research organization (CRO) drug development solutions (5). In January 2025, Lyndra Therapeutics, of Watertown, Mass., announced a strategic collaboration with Thermo Fisher leveraging the Accelerator Drug Development platform and allowing Lyndra to use Thermo Fisher’s end-to-end, advanced pharmaceutical manufacturing capabilities based out of Cincinnati, Ohio, to manufacture commercial materials at scale (6,7).

This partnership is indicative of a current trend toward custom, specialized relationships based on specific needs—in this case, Lyndra’s long-acting oral therapies including its lead investigational product, oral weekly risperdone (LYN-005)—rather than a more generalized shared-services approach. Even with that, or perhaps because of it, new concepts like the “hybrid” CDMO are emerging with more frequency. And the presence of artificial intelligence (AI) and a push for smart and/or automated technologies to pick up more and more of the work are all contributing to the shifting landscape.

®’s Drug Digest video series, Adi Kaushal, director and Technology Head of Bioavailability Enhancement at Lonza, discussed the growing preference in the market for oral solid dosage (OSD) forms, like Lyndra’s candidate, and how CDMOs can leverage their experience in performing much-sought-after expert techniques to improve OSD bioavailability to meet demand.

“CDMOs like Lonza are important partners for the smaller biotechs and emerging biotechs, and that is really driven by the experience and the expertise that CDMOs bring to the table, especially for smaller companies that may not have that experience in that space,” Kaushal said (8). “Alongside that, what comes into the picture is investment into state-of-the-art facilities and equipment, which is essential for the manufacture of a lot of these drug products. And then the final part is the regulatory knowledge and the technical knowledge that the CDMOs bring to the drug development process, partnering with the smaller biotechs. That’s really important, because the regulatory landscape is ever-changing, and it is often inherently linked to the technology in question. And so that technology expertise and regulatory expertise, from a regulatory standpoint or compliance standpoint, [are] critical to the progression of molecules into real therapies, and commercializing those.”

Trends in certain forms of drug delivery, as well as the emergence of artificial intelligence, are playing roles in evolving the nature of partnerships, but there are new types of partnerships gaining steam as well.

Featured Content

Behind the Headlines, Episode 22

ICH Q12 and Post-Approval Changes

Advancements in Biopharma Spur Innovation

What Pharma Manufacturers Need to Know

Every Pharmaceutical Manufacturing Professional Should Know