Rational Antibody Design and Engineering with Next Generation Protein Sequencing

In this webinar, you will learn:
A strategy for generating recombinant mAbs and antibody derivatives directed towards antigens involved in mitotic cell division
Methods for antibody engineering and customization, species switching, and construction of antibody fragments
How Next Generation Protein Sequencing (NGPS) works
Applications of NGPS to aid engineering and recombinant production of antibodies
Abstract
Increasing the Versatility of Recombinant Antibodies targeting mitotic epitopes
Monoclonal antibodies (mAbs) are indispensable reagents across many basic, translational, and biomedical applications from biomolecule detection to protein purification to medical diagnostics and therapeutics. However, traditional mAbs generated in animals have their drawbacks. Concerns regarding traditionally produced mAbs include reproducibility, incomplete characterization, consistent availability, and cost. In short, good mAbs are hard to find for certain targets and biological processes such as mitosis.
To maintain genomic integrity, proper segregation of chromosomes into the two daughter cells must occur. This process is powered and regulated by the kinetochores, which are structures built at the primary constriction of mitotic chromosomes that attach to spindle microtubules. The primary factor connecting the kinetochores to the microtubules is the kinetochore-associated NDC80 complex.
Advancements in protein sequencing techniques have made it possible to acquire the primary amino acid sequence of mAbs that target the NDC80 complex of the mitotic process. The invariant primary sequences were used for downstream construction and production of sequence-defined recombinant mAbs and antibody fragments, including scFvC, scFv, and Fab, all showing robust binding to the kinetochore target in mitotic cells. Moreover, this methodical approach circumvented the issues listed above, providing a straightforward and accessible strategy for generating low-cost, high-yield preparations of recombinant mAbs and antibody fragments.
Dr. Jennifer DeLuca is a Professor and Associate Chair of Graduate Studies in the Department of Biochemistry and Molecular Biology at Colorado State University. Her lab uses biochemistry and cell biology approaches to study mitosis and chromosome segregation; they are investigating why cancer cells are prone to chromosome missegregation and how that can be exploited for therapeutics. Her lab also studies how key phosphorylation and dephosphorylation events on mitotic proteins precisely regulate chromosome segregation, and has recently been developing mAb-based tools to track these post-translational events in real time. Jennifer holds a PhD in Molecular, Cellular, and Developmental Biology from the University of California in Santa Barbara. The most recent of her many awards and accolades is the 2021 American Society for Cell Biology (ASCB) Lifetime Achievement Fellow Award.
Dylan Brethour, MSc
Rapid Novor
Scientific Sales Executive
Dylan graduated from the University of Toronto with a Master’s in Laboratory Medicine and Pathobiology. He is passionate about science and new technologies, particularly about accelerating research through next generation protein sequencing. Driven by a very curious mind, Dylan thrives in problem-solving and ensuring Rapid Novor’s clients’ needs are always met.