A platform-agnostic, function first-based antibody discovery strategy using plasmid-free mammalian expression of antibodies
Ruijun Zhang, Ponraj Prabakaran, Xiaocong Yu, Brian C. Mackness, Ekaterina Boudanova, Joern Hopke, José Sancho, Jacqueline Saleh, HyunSuk Cho, Ningning Zhang, Helene Simonds-Mannes, Samuel D. Stimple, Dietmar Hoffmann, Anna Park, Partha S. Chowdhury, Sambasiva P. Rao
Abstract
transcripts could be converted to LECs, and nearly 100% of them expressed as antibodies when transfected into Expi293F cells. The concentration of IgG in the cell culture supernatants ranged from 0.05 µg/ml to 145.8 µg/ml (mean = 18.4 µg/ml). Antigen-specific binding was displayed by 78-100% of antibodies. High throughput functional screening allowed the rapid identification of several functional antibodies. In summary, we describe a plasmid-free system for cloning and expressing antibodies isolated by different approaches, in any format of choice for deep functional screening that can be applied in any research setting during antibody discovery.