Two-dimensional high-throughput on-cell screening of immunoglobulins against broad antigen repertoires

• Published in: Communications biology Vol. 7; no. 1; pp. 842 - 13
• Main Authors: Lomakin, Yakov A., Ovchinnikova, Leyla A., Terekhov, Stanislav S., Dzhelad, Samir S., Yaroshevich, Igor, Mamedov, Ilgar, Smirnova, Anastasia, Grigoreva, Tatiana, Eliseev, Igor E., Filimonova, Ioanna N., Mokrushina, Yuliana A., Abrikosova, Victoria, Rubtsova, Maria P., Kostin, Nikita N., Simonova, Maria A., Bobik, Tatiana V., Aleshenko, Natalia L., Alekhin, Alexander I., Boitsov, Vitali M., Zhang, Hongkai, Smirnov, Ivan V., Rubtsov, Yuri P., Gabibov, Alexander G.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.07.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 13/1; 13/109; 13/31; 631/154/51/1568; 631/1647/2163; 631/1647/664; 631/250/1619/40/1774; 692/699/255/2514; 82/103; Affinity; Antibodies; Antibodies, Viral - immunology; Antibody libraries; Antigens; Biomedical and Life Sciences; COVID-19 - immunology; COVID-19 - virology; Epitope mapping; Epitope Mapping - methods; High-Throughput Screening Assays - methods; Humans; Immunogenicity; Immunoglobulins; Immunoglobulins - genetics; Immunoglobulins - immunology; Life Sciences; Lymphocytes B; Monoclonal antibodies; Pathogens; Peptide Library; Peptide mapping; Phages; Proteins; SARS-CoV-2 - immunology; Severe acute respiratory syndrome coronavirus 2; Spike Glycoprotein, Coronavirus - genetics; Spike Glycoprotein, Coronavirus - immunology; Spike protein; Transcriptomes
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-024-06500-2

Identifying high-affinity antibodies in human serum is challenging due to extremely low number of circulating B cells specific to the desired antigens. Delays caused by a lack of information on the immunogenic proteins of viral origin hamper the development of therapeutic antibodies. We propose an efficient approach allowing for enrichment of high-affinity antibodies against pathogen proteins with simultaneous epitope mapping, even in the absence of structural information about the pathogenic immunogens. To screen therapeutic antibodies from blood of recovered donors, only pathogen transcriptome is required to design an antigen polypeptide library, representing pathogen proteins, exposed on the bacteriophage surface. We developed a two-dimensional screening approach enriching lentiviral immunoglobulin libraries from the convalescent or vaccinated donors against bacteriophage library expressing the overlapping set of polypeptides covering the spike protein of SARS-CoV-2. This platform is suitable for pathogen-specific immunoglobulin enrichment and allows high-throughput selection of therapeutic human antibodies. The authors present a 2D screening approach using antibody libraries against a panel of antigen fragments exposed on the phage surface, which facilitates the mapping of monoclonal antibodies to panels of diverse antigens, even with unknown immunogens.