The U.S. Food and Drug Administration (FDA) approved the first DNA recombinant protein – human insulin (Humulin) – in 1982, setting the precedent for using recombinant technologies to generate therapeutics.
From producing cutting-edge vaccines to cancer-fighting medicines, there has been rapid growth in the use of cell lines to create a growing number of biologics entering the market targeting a wide variety of therapeutic areas. The blockbuster status of many biologics and advancements in biotechnology have driven the growth of this drug development sector.
Cell line development is the linchpin of biotherapeutic production. The choices made at this stage will ultimately influence success and poor decisions will be magnified as the project progresses. Therefore at the earliest opportunity during the development of cell lines, developers should be planning ahead to consider how decisions at this stage will influence potential challenges in upstream, downstream and formulation processes.
In our downloadable whitepaper, learn about how cell line development (CLD) is the linchpin of biotherapeutic development and manufacturing and how processes can be streamlined so programs can reach IND faster.
What is CLD?
CLD refers to the process in which living cells are used to manufacture and produce therapeutic biologics or other significant proteins. The CLD process begins with small-scale suspension cultures which then progress to industrial-scale bioreactors during the manufacturing phase. By getting the processes right, the balance of high quality, high yield, and speed needed to bring a biologic to market successfully can be achieved by working with an experienced CLD services partner.
Why is CLD so important?
CLD is challenging. Establishing cell lines is a long process, with numerous phases to navigate and de-risk. Meeting the increasing global demand for biotherapeutics is also a significant challenge that needs to be met, and relies on the ability to reproducibly deliver biologics at scale. The success of moving a biologic project from lab – to industrial scale hinges on cell line expertise.
At Abzena we have seen that Monoclonal Antibodies (mAbs) and their derivatives – including Fabs and Fc fusions – have shown remarkable market growth in the past decade. The use of mAbs as therapeutic agents has become routine for the treatment of many diseases such as asthma, macular degeneration, arthritis, Crohn’s disease, and various types of cancer.
How can Abzena help develop cell lines efficiently and reduce risk?
With over 20 years of expertise and a commitment to quality excellence, we can reduce CLD timelines by >30%. Our proprietary AbZelectPRO™ platform accelerates timelines and de-risks CLD programs. Recently enhanced, AbZelectPRO™ efficiently enables the rapid delivery of high-producing cell lines up to 8g/L for complex biologic or bioconjugate development programs.
Building a Robust CLD Strategy
IND applications require a great deal of information surrounding the manufacturing processes, as well as results of pharmacology and toxicology studies. As a result, it can take up to 18 months to make the biologic and prepare and submit an application. With the right approach to CLD, timelines can be drastically shortened, however, it is important to determine at the design stage how all processes downstream the development of cell lines will be impacted and de-risked.
- Analytics and Validation: Critical quality attributes (CQA) must be
determined and monitored all the way through the CLD and development process.
- Scalability: When scaling, developers will encounter a variety of operating parameters that can affect biological attributes, such as product yield, cell growth, and quality.
- Upstream Development (USP): Selecting clones that are a good platform fit as part of the cell line development process, considering both productivity and product quality, will be critical to maximizing the value of the platform.
- Downstream Process (DSP): Selecting lead candidates with minimal heterogeneity and using the cell line development process to identify production clones that consistently express in platform USP conditions will minimize product heterogeneity. More consistent product coming out of USP will allow for the application of platform DSP methods, reducing timelines to develop a robust, high yielding process.
As well as employing a proactive approach to the development of cell lines that prevents timeline delays, building a robust CLD strategy requires the adoption of technologies that can accelerate timelines without being detrimental to product quality.
Vector Selection
When choosing a vector for the delivery of the target genes encoding the desired biologic, having a library of carefully designed, tried, and tested plasmids with a standard backbone can accelerate CLD and shorten timelines. The plasmid backbone should incorporate the key regulatory cis-elements that drive expression and a selection marker for screening.
For different types of antibody-based molecules, vectors encoding basic subunits of the molecule, e.g., a general IgG, could be mutated and edited to change the coding region based on the requirements of the project. For example, to ensure specificity towards a particular antigen, variable regions could be quickly cloned into these “template” vectors. The edits needed would ultimately depend on the project requirements.
Vector Integration
If using a mammalian cell line for expression (e.g., CHO) the method employed for gene integration should be carefully considered to select the approach that will produce a stable cell line that consistently expresses the right quality of product at suitable levels.
Fast Stable Pools
One technique that can significantly shorten timelines to IND filing is the generation of fast stable pools. Following molecule design and developability, small amounts of protein can be rapidly generated through transient expression for basic screening of a primary characteristic e.g., binding to antigen. However, transiently produced material comes with the risk of altered product quality attributes (PQA) as compared with final production clones.
For more in-depth analysis, bulk fast stable pools of a panel of potential drug candidates can quickly be generated in a matter of a few weeks. Stable pools can be generated using the production host CHO cell line through transfection and simple selection.
As well as reducing future delays, stable pools also enable development activities to begin in parallel, overlapping them to effectively shorten development timelines:
- Lead Candidate Selection: In depth characterization of candidate molecules expressed from stable pools may include in vivo efficacy and/or dose range finding studies.
- Clone Selection: Isolation of single cells from selected stable pools. Screening and selection of clones with high productivity and desired product quality.
- Process Development: Information on yields and purity, USP and DSP development design can begin, and developers can determine how well the molecule will fit existing platforms.
- Formulation Development: Initiated to determine optimum drug substance stability conditions and drug product form using the early purified material.
- Analytical Development: the purified material from the lead candidate stable pool can drive analytical development.
Understanding Regulatory Requirements
A common issue when developing and manufacturing a new biological entity (NBE) is that the standards and regulations surrounding the processes involved are often not fully established.
For regulatory compliance throughout the development of cell lines, it is critical that there is full documentation of the origin, source, and history of cell lines used, in line with ICH guidelines. All of the steps used in the generation of the parental cell line from the progenitor must be documented.
CLD: Delivering Essential Biologics
With constant innovation, the antibody-based biologics market is set to continue
its rapid expansion, with molecules entering broader therapeutic areas. These drugs will be essential in targeting diseases of growing prevalence like autoimmune diseases, ischaemic heart disease, and cancers and infectious disease.
Delivering these essential biologics to patients at speed will require developers
to employ a robust strategy from the outset, with CLD at the core of this. As the
foundation of the drug manufacturing process, CLD requires a proactive approach to determine potential risks and challenges to avoid delays. A robust CLD strategy also has the potential to accelerate timelines.
Request your copy of our Cell Line Development Whitepaper today and find out how Abzena can help accelerate your biologic or bioconjugate based program.
Discover AbZelectPRO™: DNA to RCB in 10 weeks
Benefits of AbZelectPRO™ with 2G UNic vector technology:
For more complex biologic and bioconjugate programs, we can deliver increased productivity through our AbZelectPRO™ platform. This combines our existing CHO cell line with ProteoNic’s premium expression vector technology 2G UNic.
- Optimized expression cassettes to maximize productivity from each cell
- Efficient GS selection marker compatible with current AbZelectPRO™ CHO Cell Line
- Proven track record – ProteoNic is moving towards regulatory approvals with multiple programs in the clinic and works with several large pharmaceutical firms.
- Early material from highly productive stable pools for antibodies, bi-specifics, and fusion proteins
- Higher frequency of high-producing clones offering more shots on goal- up to 8 g/L
- Highly stable clones maintaining productivity beyond 60 generations
- Quicker doubling times with a >30% reduction in cell line development timelines
- Uses single-use bioreactors to lower production costs
- Robust scalable and consistent platform process
- Further process intensification can boost titers to achieve 10g/L at scale
- Enhanced cycle times with faster growth with optimized CHO-K1 host cell line
Request a copy of our AbZelectPRO™ Info Sheet on improving cell line development efficiency by clicking the button below.
AbZelectPRO™ Info Sheet
