Antibody Light Chains: Key to Increased Monoclonal Antibody Yields in Expi293 Cells?

• Published in: Antibodies (Basel) Vol. 11; no. 2; p. 37
• Main Authors: Gong, Siqi, Gautam, Seijal, Coneglio, Joshua D, Scinto, Hanna B, Ruprecht, Ruth M
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 18.05.2022; MDPI
• Subjects: Antigenic determinants; Antigens; Chains; Chemical properties; Cloning; Communication; Epitopes; FDA approval; Genes; HIV; Human immunodeficiency virus; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoprophylaxis; lambda light-chain preference; Light; Light chains; mAb yield; Monoclonal antibodies; monoclonal antibody; Mucosa; Plasmids; Production processes; recombinant dIgA; recombinant IgG; recombinant IgM; Recombinant proteins; Structure; United States; United States > US; Switzerland; recombinant IgM; recombinant IgG; mAb yield; monoclonal antibody; recombinant dIgA; lambda light-chain preference
• ISSN: 2073-4468; 2073-4468
• DOI: 10.3390/antib11020037

When constructing isogenic recombinant IgM-IgG pairs, we discovered that μ heavy chains strongly prefer partnering with λ light chains for optimal IgM expression in transiently cotransfected Expi293 cells. When μ chains were paired with κ light chains, IgM yields were low but increased by logs-up to 20,000 X-by using λ chains instead. Switching light chains did not alter epitope specificity. For dimeric IgA2, optimal expression involved pairing with λ chains, whereas light-chain preference varied for other immunoglobulin classes. In summary, recombinant IgM production can be drastically increased by using λ chains, an important finding in the use of IgM for mucosal immunoprophylaxis.