Application of phage-displayed single chain antibodies in western blot.
A phage display single chain fragment variable library constructed on pIII protein of M13 filamentous phage was screened using B-lymphocyte stimulator and FP248 as selective molecules. After four rounds of panning, there was a remarkable enrichment in the titer of bound phages. Twenty phage clones were selected from the last round and screened by means of phage-ELISA. With the antibody phages as primary antibodies in Western blot, we developed a method for detecting the specific antigen. The dilutions of antibody phages depend on the affinity between antibody-displayed phage particles and antigens.
Ding
Yan-Li
YL
School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, China.
Liu
Mei-Yun
MY
Han
Wei
W
Yang
Sheng-Li
SL
Liu
Hui
H
Gong
Yi
Y
D000818:Animals; D000917:Antibody Formation / Q000502:physiology*; D000936:Antigen-Antibody Complex / Q000032:analysis*; D015153:Blotting, Western / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007128:Immunoglobulin Fragments / Q000276:immunology* / Q000302:isolation & purification*; D051379:Mice; D019151:Peptide Library
D000936:Antigen-Antibody Complex; D007128:Immunoglobulin Fragments; D019151:Peptide Library