Susan Haigney

Young female tech or scientist performs protein assay | Image Credit: © tilialucida - stock.adobe.com

This year marks the 10th anniversary of the International Day of Women and Girls in Science, which was recognized on Feb. 11, 2025. There has traditionally been an unbalanced gender gap in the areas of science, technology, engineering, and mathematics (STEM) that is limiting the potential of skilled workers in the pharmaceutical industry. According to the United Nations, fewer than 30% of engineering graduates are women (1). Women also face unique challenges in these fields, including a lack of opportunity in senior positions. With the demand for highly skilled personnel high, what is the bio/pharmaceutical industry doing to encourage girls to enter STEM fields and women to pursue higher level jobs in pharma?

Michelle Bridenbaker, COO, Unbiased Science, stresses the critical nature of STEM programs in developing talent and ensuring a sustainable, and relevant, workforce. “In terms of the pharma industry specifically, a strong scientific background is a solid foundation for the rigor, discipline, and critical as well as innovative thinking that modern pharma success is built on,” says Bridenbaker. “As employers, pharma companies need to actively encourage women through mentorship and leadership preparation, so that they see and realize their full potential.”

Opportunities for STEM-related jobs in the pharmaceutical industry are abundant and are continuing to grow, according to Agnes Cwienczek, director of Product Management at ArisGlobal. It is crucial to foster education in these fields to meet the growth of technology.

“While more and more pharma companies have established dedicated STEM programs and are fostering related career paths, men continue to outnumber women in this field across the globe, particularly at the higher levels,” says Cwienczek. “It should be in our collective interest to invest further in these kind of promotions—not only for purposes of equality but also to capitalize on the skills and significant contributions made by women to science and research.”

Gender ‘norms’ often inhibit girls from entering what are traditionally seen as school subjects that are ‘for boys’. Pushing back on these stereotypes is crucial, not just for girls, but also for boys. Equal access to STEM fields without labeling specific areas as ‘gender-specific’ is important for STEM programs, says Sabine Gölden, eLearning & Training Lead at MAIN5.

“Gender should never be the basis for deciding which field of study is right for you, regardless of any evolutionary advantages or disadvantages,” says Gölden. “Ultimately, someone’s path should be determined by their unique talents, passion, dedication, and interests, not by their gender. As a mother of two, I observe with great interest how societal expectations continue to shape role models,” remarks Gölden. “A clear example is toy advertisements—on many popular websites, you can filter toys by ‘Boys’ or ‘Girls,’ resulting in distinctly different suggestions. And this is just one of many examples. That’s why I strongly believe STEM programs in schools are a valuable opportunity to challenge stereotypes and give every student the chance to discover whether STEM is the right fit for them.”

To foster interest in science and engineering for girls, exposure to these fields at an early age is imperative. “STEM programs for girls in school play a crucial role in fostering interest, confidence, and skills in science, technology, engineering, and mathematics,” explains Jean Redmond, the chief operating officer at Biologit. “These programs provide early exposure to a wide range of career possibilities, helping to break down stereotypes and encourage more girls to pursue STEM fields. Given that technology and data-driven roles are often male-dominated, targeted initiatives can help bridge the gender gap and ensure that young women feel supported in pursuing these careers.”

Bridenbaker concurs. “Everything starts at school in terms of cultivating ambition and a sense of what you can achieve,” she says. “When I was at school and passionate about science, I was at times discouraged from pursuing this as a specialist subject and career on the basis that this would be a tough path for a woman and for raising a family; it was suggested instead that being a teacher was more realistic. Actively promoting STEM subjects to girls from an early age is essential to cultivate their passion, talent, and ambition, first so they have a vision and then so that they believe in themselves enough to pursue it.”

For example, a passion for science led Alex Tabbitt, quality control microbiologist at SERB Pharmaceuticals to study biology and chemistry and pursue a degree in forensic investigation. “I think there could be more done to encourage girls through school,” says Tabbitt. “The sciences can be hard and inaccessible, which could be tackled. I was always most engaged by the practical side of my studies—the experiments and the chance to see science in action. Perhaps more could be done to link what is learned in the classroom to its significance in the real world.”

Redmond points to programs like Girls Who Code, which helps girls build coding and AI skills, as gateways for women in healthcare industries. “By increasing representation in STEM fields, these initiatives help drive diversity in leadership and innovation in pharma and biotech,” Redmond says. “Additionally, mentorship, networking, and industry partnerships further support women in transitioning from education to impactful careers.”

Ensuring women have access to grants and jobs in STEM fields requires a ‘multi-faceted approach’ of financial support, mentorship, and changes in cultural norms, asserts Redmond. Having dedicated funding and education opportunities, including internships, for women can increase access to these STEM areas.

“Mentorship plays a key role, and many companies have established internal or external mentorship programs,” says Redmond. “Professional networks play a key role. I have personally benefitted from the incredible network of women from the IQVIA WIN ERG that has lasted over many years, and engaging in these groups can lead to many opportunities such as exposure to networking events, conferences, collaboration, and job opportunities.”

“The pharma industry should absolutely be investing in and promoting STEM career paths for girls,” says Bridenbaker. “It’s incumbent on them to do so. It’s about investing in a sustainable talent base, and about doing the right thing by half of the population. Not to mention accessing the many inherent leadership qualities that women bring to the table, and as drivers of technology-based process innovation.”

Gölden points to the concept of the "leaky pipeline", which describes how women are more likely to leave STEM fields at various stages of their careers, as a factor for the lack of women in these job positions. “From my perspective the leak arises mostly from the challenge of ‘working motherhood’,” says Gölden. “There are already great set-ups as maternity leave, that jobs are held, more and more flexible working models and so on, but it is not yet enough to fully enable mothers to be able to master the work-motherhood challenge.”

While Bridenbaker believes the advancement of women in STEM fields is improving, more could be done. She points to countries, such as the United Kingdom, Ireland, France, and Nordic countries, that actively protect women in the workplace by providing maternity and paternity leave and good work-life balance practices as having a head start in creating, and maintaining, STEM jobs for women. “Without good policies here, economies and employers are putting at risk a significant proportion of their workforce and future talent base, and by extension their long-term sustainability,” she says.

“Creating a supportive workplace is vital, with active promotion, by example,” says Cwienczek. “Also, ensuring a degree of flexibility and allowance for a heathy work-family balance; though, this is not relevant only for STEM fields.”

“In particular for women, a focus on supporting a return to work following maternity leave is a crucial area where women can benefit from support in their careers,” agrees Redmond. “Finally, increasing visibility of successful women in STEM provides role models and helps shift industry norms. Providing remote work, flexible hours, and parental leave options helps retain women in STEM careers.”

 for more coverage on the International Day of Women and Girls in Science.

1. UN. International Day of Women and Girls in Science, 11 February. 

https://www.un.org/en/observances/women-and-girls-in-science-day/

Articles

Investing in STEM programs for girls in an education setting and then supporting women as they excel in their professional careers is key to ensuring that vital highly skilled personnel are not left behind. 

Leaving STEM Skills on the Table

International Day of Women and Girls in Science: Research, Diversity, and AI

 anniversary of the International Day of Women and Girls in Science. 

® sat down with Jean Redmond, Chief Operating Officer at Biologit, to talk about the importance of diversity in research, the unique role women play in the field of artificial intelligence (AI), and what can be done to enrich programs to encourage girls to pursue jobs in science, technology, engineering, and mathematics (STEM).

 AI is a fast-growing technology in the pharmaceutical industry. Are there unique and specific skills and/or viewpoints women in STEM jobs bring to this area?

 I believe AI is set to transform the pharmaceutical industry, and this is already taking place in many real-life use cases. Women in STEM need to be intimately involved in the development and deployment of AI to ensure it is built ethically and representatively and in a fit-for-purpose way for problems that affect women.

Whilst AI is hugely powerful and set to transform the industry, it needs to be understood as a tool first and not a solution out of context. Women in STEM need to provide their input to understand the unique problems and ensure appropriate and impactful application. For example, AI and automation is set to transform the landscape of clinical trials, an area in which women have been historically underrepresented. There is huge scope and opportunity here for women to play a key part in.

In addition, it has long been known that data bias exists in research, and it is important that AI models that may impact healthcare are trained on relevant and diverse datasets. In addition to gender diversity, it is also important to consider racial and age diversity and different stages of life, and ensuring the datasets are comprehensive and representative. Women can play a role in advocating for the ethical development of AI.

 How can access to grants for women scientists and to jobs for women working in STEM fields be increased and/or nurtured?

 Increasing access to grants and job opportunities for women in STEM requires a multi-faceted approach that includes financial support, mentorship, and cultural shifts.

Dedicated funding or grants to support paid industry placements for students, or further education opportunities for women, can increase access to roles in STEM. More financial aid and early exposure to STEM careers through internships can encourage long-term participation. In particular for women, a focus on supporting a return to work following maternity leave is a crucial area where women can benefit from support in their careers.

Mentorship plays a key role, and many companies have established internal or external mentorship programs. Professional networks play a key role. I have personally benefitted from the incredible network of women from the IQVIA WIN ERG that has lasted over many years, and engaging in these groups can lead to many opportunities such as exposure to networking events, conferences, collaboration and job opportunities.

Finally, increasing visibility of successful women in STEM provides role models and helps shift industry norms. Providing remote work, flexible hours, and parental leave options helps retain women in STEM careers.

Jean Redmond, Chief Operating Officer at Biologit

Jean Redmond is Chief Operating Officer at Biologit, a technology company revolutionizing literature monitoring for pharmacovigilance. At Biologit, Redmond oversees operations and lead strategic growth initiatives, leveraging her background in general management, strategy, operations, and scientific research.

Videos

Pharmaceutical Technology® sat down with Jean Redmond, Chief Operating Officer at Biologit, to discuss the use of artificial intelligence in pharmaceutical research and development and the role women play in this fast-developing area.

Black female scientist looking through microscope while working in laboratory. | Image Credit: ©Drazen - stock.adobe.com

 anniversary of the International Day of Women and Girls in Science, recognized on February 11. This day puts a spotlight on gender equality in the fields of science, technology, engineering, and mathematics (STEM). Even though progress has been made with the education of girls in STEM fields, there is still a divide when it comes to representation of women in STEM industries, such as the pharmaceutical industry, despite women being half of the world’s population (1,2). According to the United Nations (UN), only 28% of engineering graduates are women, and there are still challenges for the women who do enter into STEM fields, including sexual harassment and a lack of senior positions for women (1).

A variety of pharmaceutical companies and organizations are celebrating the day in a variety of ways. Sanofi is highlighting five employees the company feels are helping to “pave the way for change in science”: Kasey Boynton, senior director, Global Operations & Partnerships, who works to increase clinical trial access in underserved communities; Amy Brennan, cluster head, Manufacturing & Supply, who is a leader in rare diseases, oncology, and immunology, and optimizes productivity across specialty care sites; Alexandra Hicks, head of Type 2 Immunology Research, who leads research in type 2 inflammation across immune-mediated diseases; Brenda Mulligan, global head of Science and R&D Communications, who works to educate on discoveries in the lab and their impact on patients; and Sandrine Samson, head of Global Vaccine Medical Affairs, New Products & Innovation, who states, "Being part of the family of scientists and using science to help the world be healthier and more equitable is the best part" (3).

Medicines Discovery Catapult (MDC) is also highlighting women in science by featuring some of their collaborators, including Kristy Smitten, leader of MetalloBio, who has won awards for the novel antimicrobial class to combat antimicrobial resistance; Helen McCarthy, CEO and co-founder of pHion, who invented a proprietary cell-targeting drug-delivery peptide; Helen Philippou, founder of LUNAC Therapeutics, who is a researcher on blood coagulation and whose company is developing first-in-class, next-generation intravenous anticoagulants; Kate Cameron, founder of Cytochroma, which manufactures multicellular hepatic and cardiac models; and Donna Finch, head of Translational and Clinical Sciences at Alchemab Therapeutics, which was awarded best-emerging biotech start-up in at the 2021 OBN Awards (4).

Research organization MRIGlobal highlighted scientists Chelsey Smith, whose research has supported development of a massively multiplexed detection (MMD) device for early threat detection, and Alison Hart, whose works have helped develop multiple steps for synthetic routes. “For more than 80 years, MRIGlobal’s scientific research contributions have been advancing knowledge and shaping the world. These are but two of our many scientists who are women whom we celebrate,” the company stated (5).

Yasmine Sherif, executive director of Education Cannot Wait, released a statement on Feb. 11, 2025, saying, “The future of the human race hangs in the balance. With science and technology far outpacing the capacity of most humans to keep up, we must arm our future scientists and future leaders with the knowledge, skills and critical thinking abilities they need to survive and thrive in the brave new world of the 21st century. To achieve these goals, we must empower an entire generation of girls in crises to receive the education, training and resources they need to improve their knowledge and skills base in [STEM]” (6). Sherif specifically pointed out the women and girls living in areas of conflict.

“It is not an easy road. For girls living on the frontlines of armed conflict, climate change and forced displacement, the thought of even attending school is elusive at best. In all, there are now nearly a quarter of a billion crisis-affected girls and boys whose right to a quality education fit for the 21

 century is interrupted by these preventable protracted crises. Girls are among the most vulnerable. Rather than studying science or learning about technology, they are exposed to forced child marriage and unwanted pregnancies without their potentials ever being achieved,” Sherif said in the statement (6).

® will be speaking with several women working in STEM fields to discuss the impact women have in the industry. Our 

, COO of Unbiased Science, is now available (7). 

United Nations. International Day of Women and Girls in Science, 11 February. 

Sanofi. Celebrating Women in Science. Feb. 11, 2025 

https://www.sanofi.com/en/magazine/our-science/celebrating-women-science

MDC. MDC Celebrates International Day of Women and Girls in Science. 

https://md.catapult.org.uk/news/mdc-celebrates-international-day-of-women-and-girls-in-science/

MRIGlobal. Using Science to Improve People’s Lives. Blog Post, 

Education Cannot Wait. International Day of Women and Girls in Science Statement by Education Cannot Wait Executive Director Yasmine Sherif. 

https://www.educationcannotwait.org/news-stories/directors-corner/brave-new-world

Haigney, S. International Day of Women and Girls in Science: Support and Empowerment. 

https://www.pharmtech.com/view/international-day-of-women-and-girls-in-science-support-and-empowerment

Feb. 11, 2025 marks the 10th anniversary of the International Day of Women and Girls in Science.

The 10th Anniversary of the International Day of Women and Girls in Science

Futuristic, AI and business woman, cyber data and connectivity, iot overlay and technology innovation. Digital transformation, tech analytics and mockup space, web dashboard and internet holographic | Image credit: © Angelo/peopleimages.com – stock.adobe.com

The International Day of Women and Girls in Science celebrates its 10th anniversary on Feb. 11, 2025. This day recognizes the challenges of gender equality in the fields of science, technology, engineering, and mathematics (STEM) that girls and women face (1). It also celebrates the progress made to combat the bias in these fields and the important contributions women have made in STEM-related industries, such as the bio/pharmaceutical industry.

One such area is the utilization of artificial intelligence (AI) in pharmaceutical development and manufacturing. Regulations in the use of AI in pharma have been rapidly advancing (2), and use of AI requires the assistance of experts in that technology (3). Women may offer unique perspectives in this developing area, specifically when it comes to a broader look at the impact of AI. “I believe AI is set to transform the pharmaceutical industry, and this is already taking place in many real-life use cases,” says Jean Redmond, chief operating officer (COO) at Biologit. “Women in STEM need to be intimately involved in the development and deployment of AI to ensure it is built ethically and representatively and in a fit-for-purpose way for problems that affect women.”

Redmond stresses that AI, and its potential use as a tool must be understood, and the perspective women bring must be heard. “Women in STEM need to provide their input to understand the unique problems and ensure appropriate and impactful application. For example, AI and automation is set to transform the landscape of clinical trials, an area in which women have been historically underrepresented. There is huge scope and opportunity here for women to play a key part,” says Redmond.

Data biases must also be addressed, continues Redmond, and AI models used in healthcare should be trained on relevant and diverse datasets. “In addition to gender diversity, it is also important to consider racial and age diversity and different stages of life, and ensuring the datasets are comprehensive and representative. Women can play a role in advocating for the ethical development of AI.”

Women’s ability to implement such technology-based change is a distinctive benefit, specifically in identifying opportunities to improve productivity, efficiency, and attention to detail, according toMichelle Bridenbaker, COO of Unbiased Science. “We instinctively perform risk assessments, for instance. In an AI context, these qualities are especially important because of the need to consider both the ethics of AI use and also the human impact of process automation. Although it sounds like a generalization, we also tend to be very open to sharing best practice, to collaborating, and to supporting others when they are unsure how to implement AI and technology effectively. Given the speed with which AI is advancing and changing, that’s vital.”

Emotional intelligence, creativity, critical thinking, and problem solving might be characteristics that women specifically contribute to the use of AI in pharma, remarks Agnes Cwienczek, director of Product Management at ArisGlobal. “While some of these are perhaps less directly relevant to defining and developing AI based solutions, they can be important to the implementation of those solutions within an organization,” says Cwienczek.

 for more coverage of the 10th anniversary of the International Day of Women and Girls in Science. Our interview with 

2. Schmitt, S. Regulations in AI and Other Digital Technologies. 

3. Thomas, F. and Haigney, S. Quality Considerations for Using AI in Bio/Pharma. 

International Day of Women and Girls in Science: Support and Empowerment

. Feb. 10, 2025. https://www.pharmtech.com/view/international-day-of-women-and-girls-in-science-support-and-empowerment

In a lead up to The International Day of Women and Girls in Science, Pharmaceutical Technology® takes a look at the unique perspective women bring to the use of artificial intelligence in the pharmaceutical industry.

Women, STEM, and AI in Pharma

 Anniversary of the International Day of Women and Girls in Science (1), which looks to highlight the challenges and progress of women in the fields of science, technology, engineering, and mathematics (STEM). 

® sat down with Michelle Bridenbaker, COO of Unbiased Science, to gain her perspective on how women can be empowered to excel in these crucial roles, as women provide unique and important perspectives.


 What are some of the benefits of STEM programs for girls in school? How do these programs translate to women building careers in the pharmaceutical industry?

Everything starts at school in terms of cultivating ambition and a sense of what you can achieve. When I was at school and passionate about science, I was at times discouraged from pursuing this as a specialist subject and career on the basis that this would be a tough path for a woman and for raising a family; it was suggested instead that being a teacher was more realistic. Actively promoting STEM subjects to girls from an early age is essential to cultivate their passion, talent, and ambition, first so they have a vision and then so that they believe in themselves enough to pursue it. STEM programs are critical not just for fostering talent from the earliest opportunity, but also for ensuring that enough bright minds come through into scientific fields, to ensure a sustainable, relevant, and representative workforce which is vital to each country’s competitive stance in the fields of medicine, pharma, medical device development, and more. In terms of the pharma industry specifically, a strong scientific background is a solid foundation for the rigour, discipline, and critical as well as innovative thinking that modern pharma success is built on. As employers, pharma companies need to actively encourage women through mentorship and leadership preparation, so that they see and realise their full potential.

 Is there sufficient investment in promoting STEM career paths for girls? Should the pharma industry invest in these types of promotions?

The situation is improving now, but there is always a lot more than could be done. The pharma industry should absolutely be investing in and promoting STEM career paths for girls. It’s incumbent on them to do so. It’s about investing in a sustainable talent base, and about doing the right thing by half of the population. Not to mention accessing the many inherent leadership qualities that women bring to the table, and as drivers of technology-based process innovation.

 Artificial intelligence (AI) is a fast-growing technology in the pharmaceutical industry. Are there unique and specific skills and/or viewpoints women in STEM jobs bring to this area?

Women excel at instigating and implementing technology-based change, because we are inherently good at identifying opportunities to hone productivity and efficiency, while also being methodical and having a close eye on the detail. We instinctively perform risk assessments, for instance. In an AI context, these qualities are especially important because of the need to consider both the ethics of AI use, and also the human impact of process automation. Although it sounds like a generalization, we also tend to be very open to sharing best practice; to collaborating; and to supporting others when they are unsure how to implement AI and technology effectively. Given the speed with which AI is advancing and changing, that’s vital.

 Which countries or regions are at the forefront of advancing women’s role in fields like pharmaceutical development and manufacturing?

I have been a leader with reach across most geographies, and I have to say I think we still have a way to go, despite having made significant gains in the past 20 years. What I can say for certain, is that countries that actively protect women in the workforce have a head start. I’m thinking about the Nordics, the UK and Ireland, and France—where there is generally strong support in the form of maternity and paternity leave, provision for a gradual return to work, and the ability to balance work with having a family. Without good policies here, economies and employers are putting at risk a significant proportion of their workforce and future talent base, and by extension their long-term sustainability.

Click the video above for a more in-depth discussion with Michelle Bridenbaker.

UN. International Day of Women and Girls in Science, 11 February. UN.org (accessed Feb. 7, 2025) 

Michelle Bridenbaker, COO of Unbiased Science

Michelle Bridenbaker owns and is COO of Unbiased Science, a trailblazing health science communications company. She is also VP of Medical Information Leaders in Europe (MILE) and serves on the scientific boards of multiple tech and device firms, contributing to cutting-edge advancements in communications, science, and medicine.

Pharmaceutical Technology® sat down with Michelle Bridenbaker, COO of Unbiased Science, ahead of International Day of Women and Girls in Science to talk about the role of women in the pharmaceutical industry.

Movement towards the intended goal. Moving forward, step by step. Labor and aspiration. Motivation, self-development. Business strategic planning, risk management. New opportunities. Achieve. | Image Credit: ©Andrii Yalanskyi – Stock.Adobe.com

The development and production of biologics can be a complex process. And the manufacture of biopharmaceuticals requires that processes be developed and validated so that product specifications can be met to create a safe and effective biologic drug. Steps can be taken at each stage of the development and manufacturing of biologics to ensure processes are optimized for the best results.

Sponsor companies often look to contract development and manufacturing organizations (CDMOs) to help with process development of these products. The following article provides advice and insight from experts and bio/pharma CDMOs on how process development can be optimized.

A scalable production system is necessary when considering how to optimize upstream processes, according to Daniel Giroux, vice-president of Biologics Development at Abzena. Typical large-scale systems range from using 500- to 2000-L bioreactors, and the scale can prevent experiments. “If you have a single, 2000-L reactor, you can’t run multiple experiments for optimization,” says Giroux. A small-scale system will be needed in which growth and production parameters of the larger-scale system can be reproduced.

After the small-scale model is created, a design-of-experiment (DoE) approach can be used to identify parameters that may impact the titer or the product quality. “You create a statistical experiment where you’re varying those parameters in combination and singly. And then you execute that experiment in parallel bioreactor systems, so a small-scale reactor where you have many of them,” Giroux explains. Statistical tools can then be used to pull data from DoEs and models to optimize input parameters to achieve maximal titers.

Process intensification is another approach Giroux points to, where higher cell densities are used at inoculation to increase the area under the cell density versus culture time curve. “You want your cell density to be high for a longer period of time,” Giroux says. “And by starting at a higher cell density, it lets you do that, and you basically get 50 to 100% more titer in an intensified fed batch process. That means, though, that you have to develop a perfusion process for your penultimate reactor, your last seed reactor, so that you can actually have enough cells to inoculate your production systems at these higher densities.”

According to Paul Mugford, director, Biologics Process Development at BIOVECTRA, media optimization is an important part of upstream processing, especially in microbial fermentation. Media should be formulated to support optimal growth and productivity of the microorganism, says Mugford, which includes optimizing carbon and nitrogen sources, trace metals, vitamins, and other supplements. “All media components, of course, have to be animal free,” says Mugford. “And we also choose to start our process development with representative materials that will be used at large scale. These are raw materials that meet all the required GMP [good manufacturing practice] requirements, so there’s no issues transferring from small to large scale with respect to materials.”

Other areas that can be optimized are process parameters such as temperature(s), pH, dissolved oxygen, and feed rates. These can be optimized using DoE, Mugford points out. Early screening of equipment is used for initial optimization with shake flasks or higher throughput microtiter plate format.

“Once the initial screening is done, we would then move into our smallest fermenters, which are the Amber250 parallel reactor system, or our Dagsip system, where each one of those systems can run eight fermentations in parallel for further optimization of media and feeding,” explains Mugford. “And our goal here is to maximize the growth and also the productivity, so we monitor the process extensively for the carbon source (glucose or glycerol), acetate, ammonia, CO

, and use [these] data for further optimization. After that, we’d move into our larger fermenters, such as the 10-L and 50-L fermentor before scale up.”

When looking at viral-vector-based gene therapy products, larger and better-quality virus yields can be the result of optimizing upstream processes. “In cell therapy, the key is to scale up to gain higher cell yields while still maintaining the cell morphology and the genetic stability of the cells, so that it’s the same at small scale and large scale,” says Bob Schrock, PhD, senior director, Global Head of Process Development, at Lonza Group AG.

“And [for] viral vectors, keeping [the cells] happy and in culture[s] at bigger scales, larger scales, maintaining that all important yield of the number of virus per cell, [and] making sure that that stays similar the bigger, the larger and larger vessels you go up to, [is important],” Schrock explains. Multiplicity of infection (MOI), plasmid ratios, transduction amounts, and when to harvest after transduction are other parameters that need to be taken into consideration for viral vectors, says Schrock.

Passage number studies are also important. “Cells can change after multiple passages, and that can introduce variability into your process,” stresses Schrock. “So, you want to make sure that the conclusions you reached within a cell passage of 50 are the same as it is at a cell passage of 100 or whatever constraints you put in there for your minimum and maximum cell passage numbers. So, you don’t want to forget to do that as well to make sure that your process can handle cells at a variety of different passage numbers.”

Optimization of process development for cell culture is dependent on the type of product being produced, according to Schrock. Scale and purity are two important factors to consider as well if the product is cell based or single cell.

To optimize a cell culture process, the parameters that need to be optimized must be defined, says Giroux, either titer, product quality, or both. In addition, assays should be in place to identify target product quality parameters. “Typically, for product quality, we’re looking at glycosylation patterns on proteins to make sure that they are correct, or in the case of a biosimilar, that they match the originator,” Giroux says. One must then vary input parameters, both individually and in combination, and then measure their effect on productivity and product quality.

When it comes to performing process studies for cell culture, Giroux says parameters that vary in DoE studies include choice of basal medium and concentrated feeds. For fed-batch processes, parameters include choice of additives and trace elements, cush as minerals, trace metals, vitamins, anti-foam and anti-clumping agents, and surfactants.

“We then also vary or look at temperature, especially shifting the temperature. So, we typically start our productions at 37 degrees, or 36 and a half. But sometimes you get better productivity in a longer culture duration if you reduce the temperature part-way into the run to perhaps 32 degrees or 34 degrees,” explains Giroux. “But we could study that the effect of pH on productivity is assessed as well [as] agitation, and then again, [at the start] cell density can have an impact, and that’s whether it be an intensified process or not. Sometimes there’s an optimal cell density even at non frame, non-intensified process.”

“So, those are the types of parameters that we’re studying, but we generally [are] studying them in these design of experiments approaches where we can vary them in combinatorial fashion and understand the effects of the parameters individually, but also in combination with the other parameters,” Giroux continues. “So, for example, if a parameter affects your cell density, you may need higher feeds. So, a parameter that you know can’t be optimized by itself [has] to be optimized as a family.”

Schrock says it’s important to close open manual process steps to reduce contamination risks and the cost of production by reducing the required grade of the GMP production clean room (i.e., grade C cleanrooms are cheaper to maintain than grade B). Process automation for equipment and perfusion is also important, says Schrock.

“One really great example of this is the Lonza Cocoon Instrument. This is both a functionally closed system as well as automated system for manufacture of autologous cell therapies. This is a really good example of doing both automation as well as closing the process. Once you have a closed process, you move to optimizing parameters … things like pH, dissolved oxygen, agitation, speeds, gasses, and even temperatures, sometimes in certain viral vector types,” Schrock concludes.

Quality considerations, such as regulatory guideline requirements and critical quality attributes, play a role in the design and optimization of fermentation processes, according to Mugford. First, one should decide on which microbial strain to use, depending on what type of product is being produced, because each biologic has unique strain requirements. According to Mugford, engineered Escherichia coli (E. Coli) strains provide optimal performance for recombinant proteins or plasmid DNA.

“Considerations in strain selection include the protein solubility, whether or not you get correct folding, the expression location, [and] if the target protein is in the cytoplasm or the periplasm,” says Mugford. “And there’s also modified strains that allow to correct for any codon bias or disulfide bond formation. So, selecting the appropriate strain is important up front.”

The design of the plasmid is also important for protein production, says Mugford, specifically selection of the promoter to tightly control expression of genes that encode proteins. Productivity can also be improved by codon optimization of the gene of interest and removing non-essential plasmid elements, according to Mugford. “And also, you may want tunable expression in cases where inclusion bodies are formed for the protein. [Y]ou may want to express the protein slower to minimize inclusion body formation.”

Plasmid DNA products have other challenges that require attention paid to strain selection, and plasmid design, according to Mugford. Plasmid stability can be enhanced, and endonuclease activity can be reduced due to the mutations E. coli strains designed for plasmids production. A high copy number plasmid is therefore used to maximize productivity. Mugford recommends the use of a fragment analyzer to assess clones in order to select the most appropriate to produce high-quality plasmid DNA.

“One of the challenges we’ve seen in the plasmid DNA area is when you have repetitive elements in the plasmid,” says Mugford. “Long poly(A) tails are quite common in plasmids destined for in-vitro transcription to make mRNA [messenger RNA]-based therapeutics like the COVID vaccines. Long poly(A) tails are prone to recombination events, and this can give some plasmid instability and shorten the length of the poly(A) tail leading to different length products. That affects the integrity of the plasmid. An efficient clone selection process for plasmid DNA can identify clones with a single, correct length polyA tail. Another important quality parameter in plasmid DNA is the super cooling, which is an indication of how much damage has been done to the plasmid.”

The next step in optimization of microbial fermentation is setting the fermentation parameters, such as media optimization, feed rates, and other setting to ensure scalability, says Mugford. Small-scale production needs to be able to transfer to large-scale parameters.

“[A]t the early development scale, our smallest fermentors are the 250-mL Amber fermentors for condition screening. These increase to fermentors in the 10 to 50-L range in process development, and [then up to] 200-L scale at the pilot stage. At the manufacturing scale, we can scale up to our 100-L and 1000-L single-use fermentors, or our 3000-L and two, 17,000 -L stainless-steel fermentors for microbial fermentation. To go from the smallest scale to the largest scale, one of the most important parameters is the oxygen transfer coefficient (kLa). Our chemical engineers have developed models to scale up and down while reproducing the same growth rates, and providing consistent results across scales.”

Mugford recommends single-use fermenters for their ability to quickly transition between products and reducing cleaning and testing. He also points to heat transfer as an important parameter for scale up, specifically during aggressive microbial growth. “With rapid growth in E. coli, there’s a lot of heat produced. [U]nderstanding the oxygen transfer rate for the fermenters and the cooling capacities is important when moving from small to large scale,” Mugford says. “[T]here are a lot of scale-independent parameters that can also be evaluated directly using DoE at the process development [stage].”

One way to harvest cells in a bioreactor, according to Giroux, is to use depth filtration, which involves a multi-stage depth filter followed by membrane filters that remove the cells. Large surface areas are required; therefore, producing a lot of waste. “Especially at the large scale, they’re troublesome to install and remove and so on. So, the setup time is difficult to remove,” Giroux warns. He points to continuous centrifugation as a better approach, which often uses a single-use disk stack centrifuge to extract the cells and dense components from the media.

“[We] don’t lose very much of the liquid itself [with continuous centrifugation]. So, you get a pretty thick slurry of cells that comes out. It might be 80% cell mass and about 20% media,” says Giroux. “For example, if you had 8% packed cell volume in your reactor, you could basically remove only 2%; you’d remove your 8% cells and then 2% of the media. So, your yield would actually be very high. You generally follow that with a very small area depth filter, sometimes even a specialized depth filter that’s designed to remove any additional cells. These filters can be slightly charged, and they help to remove debris as well.”

Choosing the correct scale-down equipment is important at process scale so that representative material is generated for filtration, inclusion body refolding, and downstream processing steps, says Mugford. For harvesting recombinant protein in E. coli, disc stack centrifuges are often used. A key parameter used in centrifuge scaling and performance evaluation is the Q/Sigma factor, which represents the flowrate (Q) and the sigma factor (Σ), according to Mugford. “This represents the relationship between the flow rate Q to the sigma factor, which is the equivalent settling area for a given centrifuge,” he says. “It’s a fixed number for each type of centrifuge, from small to large scale, and we find this is the most reliable parameter for scaling the centrifuge harvest.”

“To optimize this step, you would typically look at the percent solids in the input, the flow rate and discharge interval, and the bowl safety factor,” Mugford explains. “Those are important parameters and measurements. And you would monitor the turbidity of the centrate as a key metric of how well the clarification is going. The percent solids is a key metric if they will be carried forward, as in cases where the product is located within the cells.”

Because plasmid DNA is sensitive to shear, it requires a different approach for harvesting. During the discharges of the centrifuge, some damage can occur to the plasmid DNA, resulting in a reduced percentage supercoiling, stresses Mugford. He points to tangential flow filtration (TFF) as a better approach for plasmid DNA harvesting. Mugford says that optimizing the TFF requires selecting the appropriate cassette type (the micron/molecular weight size of the cassette).

“Typically, you would use an open channel cassette. You need to determine the loading, so liters per meter squared for the cassette and the flux rate,” says Mugford. “So, an advantage of the TFF harvest is that the cells can be washed during filtration, so this removes any spent media and some other impurities, which gives you a cleaner product into downstream processing. After this, you would typically go into clarification with a series of depth filters followed by cell lysis. For the lysis of cells containing proteins, you would typically use a high-pressure homogenization. Because plasmid DNA is shear sensitive, we developed a different method [in collaboration with Sartorius], using a chemical lysis in a continuous flow reactor to give a mild lysis and preserve quality through the midstream steps, allowing us to extract the plasmid DNA without doing any damage to it.”

The complex nature of biologics development and product offers a variety of ways in which companies can maximize their efforts and produce safe and effective medications. CDMOs and other contract organizations can provide sponsor companies with the expertise and infrastructure to streamline these complex processes. Planning ahead and using the correct parameters and approaches can ensure a cost-effective process.

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Vol. 49, No. 1
January/February 2025
Pages: 31–33

When referring to this article, please cite it as Haigney, S. Steps to Process Development Optimization. 

The optimization of processes for the development and manufacture of biopharmaceuticals is pivotal for producing quality products. 