Non-viral adeno-associated virus-based platform for stable expression of antibody combination therapeutics Non-viral adeno-associated virus-based platform for stable expression of antibody combination therapeutics

• Published in: mAbs Vol. 6; no. 4; pp. 957 - 967
• Main Authors: Wilmes, Gwendolyn M, Carey, Kimberly L, Hicks, Stuart W, Russell, Hugh H, Stevenson, Jesse A, Kocjan, Paulina, Lutz, Stephen R, Quesenberry, Rachel S, Shulga-Morskoy, Sergey V, Lewis, Megan E, Clark, Ethan, Medik, Violetta, Cooper, Anthony B, Reczek, Elizabeth E
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.07.2014; Landes Bioscience
• Subjects: adeno-associated virus; antibody cocktails; antibody combination therapeutics; antibody mixtures; biologics expression; Cell Line; Chromatography, Ion Exchange - methods; Coculture Techniques; Dependovirus; Drug Therapy, Combination; Gene Expression; Humans; monoclonal antibodies; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Single-Chain Antibodies - biosynthesis; Single-Chain Antibodies - genetics; Single-Chain Antibodies - isolation & purification; biologics expression; adeno-associated virus; antibody mixtures; antibody combination therapeutics; antibody cocktails; monoclonal antibodies
• ISSN: 1942-0862; 1942-0870; 1942-0870; 1942-0862
• DOI: 10.4161/mabs.28917

Antibody combination therapeutics (ACTs) are polyvalent biopharmaceuticals that are uniquely suited for the control of complex diseases, including antibiotic resistant infectious diseases, autoimmune disorders and cancers. However, ACTs also represent a distinct manufacturing challenge because the independent manufacture and subsequent mixing of monoclonal antibodies quickly becomes cost prohibitive as more complex mixtures are envisioned. We have developed a virus-free recombinant protein expression platform based on adeno-associated viral (AAV) elements that is capable of rapid and consistent production of complex antibody mixtures in a single batch format. Using both multiplexed immunoassays and cation exchange (CIEX) chromatography, cell culture supernatants generated using our system were assessed for stability of expression and ratios of the component antibodies over time. Cultures expressing combinations of three to ten antibodies maintained consistent expression levels and stable ratios of component antibodies for at least 60 days. Cultures showed remarkable reproducibility following cell banking, and AAV-based cultures showed higher stability and productivity than non-AAV based cultures. Therefore, this non-viral AAV-based expression platform represents a predictable, reproducible, quick and cost effective method to manufacture or quickly produce for preclinical testing recombinant antibody combination therapies and other recombinant protein mixtures.