Gyrolab Spin Blog

Development of high-affinity binding interactions and utilization of the appropriate tools for affinity determination are the ultimate goals of antibody therapeutic R&D. The expansion of protein engineering methods has provided powerful techniques for affinity maturation of biotherapeutics to accomplish at least part of this goal. In particular, the recent use of phage display methods for antibody affinity evolution have produced high affinity antibodies in the picomolar dissociation constant (K_D) range, also with high specificity, such as the TNF-α inhibitor Humira^® (AbbVie Inc.)^1,2

Vaccine discovery and development

Vaccines have become key weapons in the fight against infectious disease and as immunotherapies for other conditions including certain cancers and Alzheimer’s disease. Immunoassays in vaccine development play a key role and meeting ever more pressing deadlines means that efficient and flexible platforms are at a premium, especially in times like these, with the COVID-19 pandemic pressing vaccine R&D to the limit. In this two-part series we will be looking at how immunoassays can contribute to vaccine R&D and production.

Vaccine bioprocess development and production

Only clean drinking water can match vaccination in its ability to save millions of lives every year. In the case of the ongoing COVID-19 pandemic, the availability of a vaccine is considered a prerequisite for a return to normal life worldwide. But transforming antigens into robust vaccine products is a lengthy process. In the second article of this two-part series we will look at some examples of how immunoassays can play a key role in delivering reliable vaccine bioassay data to support efficient bioprocess development and production to ensure that vaccine production lots meet targets, time and time again.

Getting a meaningful result

Many good points were raised when it comes to impurity testing at the latest Gyrolab User Seminar, held in Milan in June. This included a roundtable discussion on a subject that has been a stumbling block for many – how to measure Host Cell Protein (HCP) levels in a meaningful way.

Target-specific reagents and immunoassay formats smoothen validation and transfer between CMC, preclinical, and clinical applications, and also to a CRO

Robust, accurate, precise, and time effective ligand binding assay platforms are crucial for the development of advanced bioassays for analyzing therapeutic antibody mixtures due to the inherent complexity of these drugs. This is why Symphogen A/S, Denmark chose Gyrolab system to develop assays to support chemistry, manufacturing, and controls (CMC) release and stability programs, and pharmacokinetics preclinical and clinical studies on Sym015, a novel anti-cancer biotherapeutic comprising two humanized monoclonal antibodies (mAbs).

It’s all about maintaining equilibrium

Immunoassays must be able to generate accurate and precise quantitative measurements of analytes such as drug candidates and their targets, and many studies also demand the accurate determination of levels of free or bound drug, or target. Achieving this is a challenge and success depends on many factors, including taking steps to maintain the equilibrium formed in vivo between various molecular forms of the analyte. This can be achieved with care in sample preparation, and by using an immunoassay platform that requires minimal sample dilution, and delivers results quickly before the equilibrium can shift.

With the need to increase analytical throughput in biotherapeutic R&D and cell and gene therapy, multiplexing assays can appear to be an obvious approach. There are certainly potential advantages, but also disadvantages and considerable challenges involved in multiplexing immunoassays – assays must be carefully matched to function in the same buffer and in the same analytical range. An alternative approach – to miniaturize singleplex ELISAs and run them in parallel – may be even more attractive.

Immunoassays have become invaluable in the generation of data for preclinical and clinical studies, process development/optimization and manufacture of biotherapeutics, and also for new applications such as determining viral titer in cell and gene therapy. Having confidence in the data based on the accurate and precise determination of analytes in complex matrices means overcoming many challenges. These can include interference that reduces robustness, selectivity and specificity, the poor performance of cross-reacting antibody reagents, and time-consuming methods that threaten time plans. Such challenges can be overcome by choosing the right reagents, using the right approach on the right immunoassay platform to deliver reliable data as quickly as possible.

With R&D returns for large cap biopharma companies falling to the lowest level in nine years (Deloitte, 1), the escalating cost of bringing medicines to market means that the biopharma R&D process must be examined in detail. One important factor is the development and validation of immunoassays that can deliver high quality data efficiently and in a timely manner. Speed and quality are intertwined, and while manual ELISA has been a major workhorse in biopharma R&D, automation is a key factor in speeding up the generation of high quality data with minimum hands-on time, minimum need for re-runs, and using readily validated assays that meet regulatory requirements.

Careful platform choice generates data with higher quality to support DOE

Harnessing the power of cell and gene therapy involves a large range of analytical methods that can deliver robust results to support rapid data driven decision-making. One assay development team has used immunoassays to track a transgene IgG product. Their choice of immunoassay platform enabled them to generate high quality data that supported Design of Experiments to quickly optimize cell culture.