Biopanning of antibody-phage clones using immunoplates coated with gel slices of electrophoresis: immunogel-biopanning.
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.
Hamasaki
Takayuki
T
Department of Bioengineering, Faculty of Engineering, Kagoshima University, 1-21-40 Korimoto, Kagoshima 890-0065, Japan.
Uchida
Shunsuke
S
Yoshihara
Tomoki
T
Hashiguchi
Shuhei
S
Ito
Yuji
Y
Sugimura
Kazuhisa
K
D000906:Antibodies / Q000737:chemistry*; D000918:Antibody Specificity; D001435:Bacteriophages / Q000737:chemistry*; D003001:Cloning, Molecular; D003114:Colony-Forming Units Assay; D004247:DNA / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007120:Immunochemistry / Q000379:methods*; D020382:Interleukin-18 / Q000032:analysis* / Q000096:biosynthesis; D053727:Interleukin-18 Receptor alpha Subunit / Q000378:metabolism; D011487:Protein Conformation; D017422:Sequence Analysis, DNA
D000906:Antibodies; D020382:Interleukin-18; D053727:Interleukin-18 Receptor alpha Subunit; D004247:DNA