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Article

BLI-Based Functional Assay in Phage Display Benefits the Development of a PD-L1-Targeting Therapeutic Antibody

by 1,2,†, 1,2,†, 1, 1, 2, 1,* and 1,*
1
Scripps Korea Antibody Institute, KNU Chuncheon Campus, Chuncheon, Gangwon 200-701, Korea
2
Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon, Gangwon 200-701, Korea
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 9 April 2020 / Revised: 9 June 2020 / Accepted: 24 June 2020 / Published: 25 June 2020
The therapeutic functionality of the antibodies from phage display is verified after an initial screening. Several immunological assays such as ELISA, flow cytometry, the western blot, and surface plasmon resonance (SPR) assay are commonly used; the IgG-format antibody is usually preferred to verify the functionality of antibodies, which need elaborative mammalian expression and purification work. Here, we describe a biolayer interferometry (BLI)-based assay that can evaluate the inhibitory functions of antibodies at an earlier stage of screening. To develop a PD-L1-targeting antibody from phage display, we applied the BLI assay to the initial scFv antibody screening, in addition to common ELISA and fluorescence-activated cell sorting (FACS) assays, which showed high advantages and relevance with the in vitro cell-based PD-1/PD-L1 inhibition assay. The same assays for IgG-format antibodies showed high efficiency of the BLI assay in the functional characterization of antibodies, and one candidate selected from the BLI assay resulted in highly efficacious antitumor activity in an in vivo syngeneic mouse study. The BLI assay was also beneficial when searching for antibodies with diverse epitopes. These results demonstrated that the BLI-based inhibition assay is an excellent technique for high-throughput scFv antibody screening in earlier stages and can make phage-display antibody screening more efficient to develop therapeutic candidates. View Full-Text
Keywords: phage display; biolayer interferometry (BLI); scFv; high-throughput screening; antibody; PD-L1 phage display; biolayer interferometry (BLI); scFv; high-throughput screening; antibody; PD-L1
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MDPI and ACS Style

Choi, J.R.; Kim, M.J.; Tae, N.; Wi, T.M.; Kim, S.-H.; Lee, E.S.; Kim, D.H. BLI-Based Functional Assay in Phage Display Benefits the Development of a PD-L1-Targeting Therapeutic Antibody. Viruses 2020, 12, 684. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060684

AMA Style

Choi JR, Kim MJ, Tae N, Wi TM, Kim S-H, Lee ES, Kim DH. BLI-Based Functional Assay in Phage Display Benefits the Development of a PD-L1-Targeting Therapeutic Antibody. Viruses. 2020; 12(6):684. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060684

Chicago/Turabian Style

Choi, Jong R., Min J. Kim, Nara Tae, Tae M. Wi, Se-Ho Kim, Eung S. Lee, and Dae H. Kim 2020. "BLI-Based Functional Assay in Phage Display Benefits the Development of a PD-L1-Targeting Therapeutic Antibody" Viruses 12, no. 6: 684. https://0-doi-org.brum.beds.ac.uk/10.3390/v12060684

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