TO STUDY INTERACTIONS BETWEEN MONOCLONAL ANTIBODY AND CHO HOST CELL PROTEINS TO DESIGN THE WASH PROCESS FOR MAB PURIFICATION

Abstract

Today, recombinant protein therapies represent a substantial focus of the pharmaceutical industry. Most therapeutic proteins are produced in host cells of non-human origin, including Escherichia coli, yeast, and various mammalian cell lines e.g. Chinese Hamster Ovary Cell Lines. A major focus of all therapeutic protein purification process is to reduce components of host organism, including host cell proteins (HCPs), to levels considered as adequate in the final formulated drug product. HCPs can pose potential safety risks for patients, including immune reactions, decreased product stability, adjuvant activity, and (theoretically) protein-specific biological activity.(Schenauer, Flynn, & Goetze, 2012) With rapidly growing cases and increased number of cancer, autoimmune diseases, Alzheimer’s disease has become the most common death-causing diseases worldwide. Recent studies indicate that mAb is effective in treatment of these diseases and with limited number of treatment options people need to rely on these medicines. Thus, the purity of these medicines becomes an important factor. If these medicines are not pure the HCP might induce antigenic reactions in the patient also these HCPs if proteolytic may degrade the desired amount of mAb to be effective as dose, thus making the medicine ineffective over a period. Thus here, we report results of the studies relating to the most interactive HCPs which isolated along with mAb and studied their interactions to design the wash process accordingly. Using the proteome of Chinese Hamster, the hotspot for the proteins were found using Aggrescan. Structure prediction was done using threading software’s Bhageerath, Pyre 2, Multicom-Raptor-X, Robetta and I-TASSER. The model generated was further validated by Independent servers Prochek, Verify-3d, Errat and with meta servers such as Protsav and SAVES. The models were refined using 3D refine and galaxy refine. The best models were docked with mAb using cluspro to identify the interactions between HCP and mAb then find the amino acid interaction profile using PDB-Sum and further work on developing modified wash process to break these bonds and obtain pure mAb to ensure effective treatment by functionally characterizing the proteins.