Biopanning of antibody-phage clones using immunoplates coated with gel slices of electrophoresis: immunogel-biopanning

Takayuki Hamasaki; Shunsuke Uchida; Tomoki Yoshihara; Shuhei Hashiguchi; Yuji Ito; Kazuhisa Sugimura

doi:10.1248/bpb.30.1361

Biopanning of antibody-phage clones using immunoplates coated with gel slices of electrophoresis: immunogel-biopanning

Biol Pharm Bull. 2007 Aug;30(8):1361-4. doi: 10.1248/bpb.30.1361.

Authors

Takayuki Hamasaki¹, Shunsuke Uchida, Tomoki Yoshihara, Shuhei Hashiguchi, Yuji Ito, Kazuhisa Sugimura

Affiliation

¹ Department of Bioengineering, Faculty of Engineering, Kagoshima University, 1-21-40 Korimoto, Kagoshima 890-0065, Japan.

PMID: 17666785
DOI: 10.1248/bpb.30.1361

Abstract

Biopanning of a phage library using a Western blotting membrane is difficult because of high background binding. We propose a reliable biopanning method, namely, immunogel-biopanning, which is performed using immunoplates coated with a molecular species fractionated from a crude sample by native polyacrylamide gel electrophoresis (PAGE). The efficacy of this method was determined in model experiments using a human interleukin-18 (IL-18)-specific single chain Fv (scFv) phage clone.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies / chemistry*
Antibody Specificity
Bacteriophages / chemistry*
Cloning, Molecular
Colony-Forming Units Assay
DNA / analysis
DNA / genetics
Electrophoresis, Polyacrylamide Gel / methods*
Enzyme-Linked Immunosorbent Assay
Humans
Immunochemistry / methods*
Interleukin-18 / analysis*
Interleukin-18 / biosynthesis
Interleukin-18 Receptor alpha Subunit / metabolism
Protein Conformation
Sequence Analysis, DNA

Substances

Antibodies
Interleukin-18
Interleukin-18 Receptor alpha Subunit
DNA