A New Method to Characterize Conformation-Specific Antibody by a Combination of Agarose Native Gel Electrophoresis and Contact Blotting
Teruo Akuta,
Toshiaki Maruyama,
Chiaki Sakuma,
Masataka Nakagawa,
Yui Tomioka,
Kevin Entzminger,
Jonathan K. Fleming,
Ryo Sato,
Takashi Shibata,
Yasunori Kurosawa,
C. J. Okumura,
Tsutomu Arakawa
Affiliations
Teruo Akuta
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Toshiaki Maruyama
Abwiz Bio Inc., 9823 Pacific Heights Blvd., Suite J, San Diego, CA 92121, USA
Chiaki Sakuma
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Masataka Nakagawa
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Yui Tomioka
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Kevin Entzminger
Abwiz Bio Inc., 9823 Pacific Heights Blvd., Suite J, San Diego, CA 92121, USA
Jonathan K. Fleming
Abwiz Bio Inc., 9823 Pacific Heights Blvd., Suite J, San Diego, CA 92121, USA
Ryo Sato
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Takashi Shibata
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
Yasunori Kurosawa
Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan
C. J. Okumura
Abwiz Bio Inc., 9823 Pacific Heights Blvd., Suite J, San Diego, CA 92121, USA
Tsutomu Arakawa
Alliance Protein Laboratories, 13380 Pantera Rd, San Diego, CA 92130, USA
In this study, we review the agarose native gel electrophoresis that separates proteins and macromolecular complexes in their native state and transfer of the separated proteins from the agarose gel to membranes by contact blotting which retains the native state of these structures. Green fluorescent protein showed functional state both on agarose gel and blotted membrane. Based on the combined procedures, we discovered conformation-specific monoclonal antibodies against PLXDC2 and SARS-CoV-2 spike protein.