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Updated: May 26, 2026

Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique
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Published on: July 19, 2019

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

Youji Shimazaki1, Yoshinori Kohno

  • 1Graduate School of Science and Engineering (Science Section) and Venture Business Laboratory, Ehime University, Matsuyama City, Japan. shimazaki.yoji.my@ehime-u.ac.jp

Analytical Biochemistry
|January 10, 2012
PubMed

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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