Class-Switching of B Lymphocytes by DNA and Cell Immunization for Stereospecific Monoclonal Antibodies against Native GPCR

• Published in: Immuno 2021-12, Vol.1 (4), p.432-441
• Main Authors: Isozaki, Yushi, Tsumoto, Kanta, Tomita, Masahiro
• Format: Article
• Language: English
• Subjects: Antigens; B-cell receptor (BCR); class-switching; DNA immunization; G protein-coupled receptor (GPCR); Genes; Lymphocytes; Monoclonal antibodies; Multiple myeloma; native antigen; stereospecific monoclonal antibody; Vectors (Biology)
• ISSN: 2673-5601; 2673-5601
• DOI: 10.3390/immuno1040031

To develop efficient applications of monoclonal antibodies for therapeutic purposes, stereospecific recognition of the target antigens is needed. DNA immunization is one of the best methods for sensitizing B lymphocytes that can produce conformation-specific antibodies. Here we verified the class-switching of monoclonal antibodies by DNA immunization followed by cell immunization for the generation of stereospecific monoclonal antibodies against native G protein-coupled receptor (GPCR) using the optimized stereospecific targeting (SST) technique. This technology facilitates the efficient selection of sensitized B lymphocytes through specific interaction with the intact antigen via B-cell receptors (BCRs). We demonstrate that multiple DNA immunizations followed by a single cell immunization in combination with a longer sensitization period (three to four months) are an appropriate sensitizing strategy for the generation of IgG-type stereospecific monoclonal antibodies by class-switching, and the characteristics of antibody production could be transferred to hybridoma cells provided by the optimized SST technique.