Identification of phage-displayed peptides which bind to the human HnRNPA1 protein-specific monoclonal antibody.
We have screened a peptide phage display library to examine if monoclonal antibody-binding phages could be isolated from the library and thereby predict the antigenic epitopes of the antibodies from the isolated phages. The library was screened for high-avidity binding to monoclonal antibodies by an affinity purification technique called biopanning. Among the monoclonal antibodies examined, the human hnRNPA1 protein-specific monoclonal antibody 9H10 showed selective binding of phages. After two rounds of the biopanning, twelve clones of high-avidity-binding phages were chosen and their inserts were sequenced. Nucleotide sequence comparison of the 12 clones showed that there were 5 different species, with two species containing four members, implying that they were predominantly selected by the biopanning. The amino acid sequences of the inserts of the 12 clones were compared with that of the human hnRNPA1 protein in order to find the putative epitope of the human hnRNPA1 protein for 9H10. The C-terminal region of the human hnRNPA1 protein shows significant homology with the peptide sequences of the selected phage clones. These results show that this peptide phage display library can be useful in defining the epitope of some monoclonal antibodies.
Kim
J H
JH
Graduate School of Biotechnology, Korea University, Seoul.
Lee
J H
JH
Kim
J
J
Kim
J K
JK
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000378:metabolism; D017104:Bacteriophage M13 / Q000378:metabolism; D018604:Epitope Mapping / Q000379:methods*; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism; D012261:Ribonucleoproteins / Q000276:immunology* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D019151:Peptide Library; D010455:Peptides; D012261:Ribonucleoproteins