Targeting antigen-specific receptors on B lymphocytes to generate high yields of specific monoclonal antibodies directed against biologically active lower antigenic peptides within presenilin 1.
We describe a targeting technique that selects antigen-specific receptors on B lymphocytes using antigen driven selective production of monoclonal antibodies which are directed against functional peptide sequences within the presenilin 1 molecule that is believed to be related to the early-onset of familial Alzheimer's disease. Three different peptide sequences of presenilin 1 were constructed, one including the region around the amino acid position 300, where the putative cleavage site exists and the other two present in the N- and C-terminal regions of that site. The efficiency in production of the desired monoclonal antibodies was at least 5-40-fold that obtained with the poly(ethylene glycol) (PEG)-mediated method. In addition, monoclonal antibodies directed against each of the peptide sequence displayed a high specificity for the corresponding peptide, in contrast to the lack of success using the PEG method. Also, the selection of surface immunoglobulin receptors on B lymphocytes by the peptides of interest was confirmed by immunofluorescent analysis. Here we demonstrate that targeting B lymphocytes results in the successful and efficient production of highly specific monoclonal antibodies against the lower antigenic peptide sequences.
Tomita
M
M
Department of Chemistry for Materials, Faculty of Engineering, Mie University, 1515 Kamihama-cho, Tsu, 514-8507, Mie, Japan. tomita@chem.mie-u.ac.jp
Sugi
H
H
Ozawa
K
K
Tsong
T Y
TY
Yoshimura
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D000941:Antigens; D001360:Avidin; D001402:B-Lymphocytes / Q000276:immunology; D003432:Cross-Linking Reagents; D006801:Humans; D007118:Immunoassay; D007158:Immunologic Techniques; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D053764:Presenilin-1; D011947:Receptors, Antigen, B-Cell / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; C508853:PSEN1 protein, human; D010455:Peptides; D053764:Presenilin-1; D011947:Receptors, Antigen, B-Cell; D001360:Avidin