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Successive analysis of antigen trapping and enzymatic digestion on membrane-immobilized avidin

Authors
Publisher
Elsevier Inc.
Publication Date
Volume
422
Issue
1
Identifiers
DOI: 10.1016/j.ab.2011.12.023
Keywords
  • Acth
  • Biotinylated Antibody
  • Biotinylated Trypsin
  • Maldi–Tof Ms
  • Immunoaffinity
  • Nondenaturing Electrophoresis

Abstract

Abstract 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.

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