Successive analysis of antigen trapping and enzymatic digestion on membrane-immobilized avidin.
Avidin from egg white was migrated toward a cathode of nondenaturing electrophoresis and then immobilized on a polyvinylidene difluoride membrane. Adrenocorticotropic hormone (ACTH) was specifically captured after the biotinylated anti-ACTH antibody was bound to the membrane-immobilized avidin, and the captured ACTH was digested by the biotinylated trypsin on the membrane after extraction. The digested polypeptides from the ACTH were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). These results indicate that target substances can be specifically trapped and digested on membrane-immobilized avidin.
Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Matsuyama City, Japan. shimazaki.yoji.my@ehime-u.ac.jp
Youji
Y
Shimazaki
Yoshinori
Y
Kohno
D000324:Adrenocorticotropic Hormone / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D000941:Antigens / Q000032:analysis* / Q000737:chemistry; D001360:Avidin / Q000032:analysis / Q000737:chemistry*; D004529:Egg White / Q000737:chemistry; D004566:Electrodes; D004789:Enzyme Activation; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry*; D007163:Immunosorbent Techniques; D008566:Membranes / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry; D059748:Proteolysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013194:Staining and Labeling; D014357:Trypsin / Q000737:chemistry; D005465:Fluorocarbon Polymers
D000324:Adrenocorticotropic Hormone; D000906:Antibodies; D000936:Antigen-Antibody Complex; D000941:Antigens; D001360:Avidin; D055767:Immobilized Proteins; D010447:Peptide Hydrolases; D011145:Polyvinyls; D014357:Trypsin; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers