Isolation of Common Light Chain Antibodies from Immunized Chickens Using Yeast Biopanning and Fluorescence‐Activated Cell Sorting. Issue 3 (8th October 2020)

• Record Type: Journal Article
• Authors: Bogen, Jan P.; Storka, Juliana; Yanakieva, Desislava; Fiebig, David; Grzeschik, Julius; Hock, Björn; Kolmar, Harald
• Is Part Of: Biotechnology journal. Volume 16:Issue 3(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-10-08
• Subjects: antibody discovery -- cell panning -- common light chain -- fluorescence‐activated cell sorting -- yeast display; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.202000240
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract: The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors. Abstract : A common light chain yeast surface display library derived from immunized chickens is generated and high affine antibody variants are isolated using yeast biopanning in combination with fluorescence‐activated cell sorting. Resulting antibodies address a broad epitope space and exhibit favorable aggregation propensities and thermal stabilities while sharing a single light chain.