Human monoclonal antibodies to viral peptides.
A new approach to the developing of human monoclonal antibodies to viral peptides is described. The method is based on the positive selection of B cells specific to viral peptides from normal human tissues. To isolate B cells bearing immunoglobulin receptors specific to viral epitopes magnetic beads coated by viral peptides were used. Human B lymphocytes specific to the neutralizing epitope of gp120 of human immunodeficiency virus and to the major epitope of Epstein-Barr nuclear antigen-I were isolated from freshly removed human tonsils. Preselected cells were transformed by Epstein-Barr virus and cultivated on irradiated human embryonic lung fibroblasts in RPMI 1640-OptiMEM medium with 10% fetal calf serum and all necessary supplements. The cultures of B cells producing antibodies to viral epitopes used for selection were obtained. Two cultures secreting IgM antibodies to the peptide MN-24 (neutralizing epitope of immunodeficiency virus) and to the peptide p107 of Epstein-Barr nuclear antigen-1 were expanded, and the specificity of the antibodies and antibody-producing cells was analyzed. The method of positive selection of B cells specific to viral peptides may be used for the preparation of human monoclonal antibodies to viral antigens.
Sidorova
E
E
Laboratory of Immunoglobulin Biosynthesis, Russian Academy of Medical Sciences, Moscow, Russia.
Borisova
T
T
Lyakisheva
L
L
Rosen
A
A
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D016175:B-Lymphocyte Subsets / Q000276:immunology*; D002461:Cell Line, Transformed; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D000939:Epitopes / Q000276:immunology; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology; D005347:Fibroblasts; D015483:HIV Antibodies / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology / Q000502:physiology; D006801:Humans; D008863:Microspheres; D008969:Molecular Sequence Data; D014066:Palatine Tonsil / Q000166:cytology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D019309:Epstein-Barr Virus Nuclear Antigens; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D010446:Peptide Fragments