Dual generation of stereo- and linear-specific monoclonal antibodies through B-cell receptors by DNA and cell immunization for therapeutic applications

• Published in: Biochimica et biophysica acta. General subjects Vol. 1869; no. 8; p. 130822
• Main Authors: Miyamae, Chiho, Isozaki, Yushi, Tsumoto, Kanta, Tomita, Masahiro
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2025
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - immunology; B-cell receptor (BCR); B-Lymphocytes - immunology; Cell Line, Tumor; DNA - immunology; DNA and cell immunization; Dual generation; Humans; Hybridomas - immunology; Immunization - methods; Intact antigen; Mice; Receptors, Antigen, B-Cell - immunology; Stereospecific monoclonal antibody; Therapeutic application; Stereospecific monoclonal antibody; Therapeutic application; DNA and cell immunization; B-cell receptor (BCR); Dual generation; Intact antigen
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2025.130822

The optimized stereospecific targeting (SST) technique features selective generation of conformation-specific monoclonal antibodies against membranous proteins with high specificity after DNA and cell immunization. This technology consists of two critical steps, which are specific selection of sensitized B lymphocytes by antigen-expressing myeloma cells through B-cell receptors (BCRs) and selective fusion of B cell-myeloma cell complexes by electrical pulses to produce hybridoma cells secreting stereospecific monoclonal antibodies. Here we were able to verify the critical step for the selection of B lymphocytes by intact antigen-expressing myeloma cells by a double-label immunofluorescence analysis. Interestingly, the cell complex was a single attachment. Furthermore, we newly found the new progress that the optimized SST technique offered dual production of anti-intact and anti-linear specific monoclonal antibodies against a human ephrin type-A receptor 2 (hEphA2). The optimized SST technique may be useful for producing not only stereospecific monoclonal antibodies, but also primary-specific monoclonal antibodies based on the selection of sensitized B lymphocytes by the target intact antigen through BCRs. It would elicit more advanced medical applications by generating dual monoclonal antibodies against the intact antigen. [Display omitted] •Dual generation of stereo- and linear-specific antibodies through B-cell receptors.•Selection of B lymphocytes by intact antigen after DNA and cell immunization.•Isotypes would determine the specificity of monoclonal antibodies.•B-cell receptors recognize stereo- and linear-specific structures of the antigen.•B cell and myeloma cell complex exhibits a single attachment in a 1:1 ratio.