[Expression of bursin receptor on membranes of hybridoma cell and identification of binding peptide of bursin from 12-merrandom phage display peptide library].
OBJECTIVE
Bursin (BS) could greatly promote the production of monoclonal antibody in hybridoma cell. We studied the interaction between BS and hybridoma cell.


METHODS
Fluorescence microscopy, Laser Scanning Confocal Microscope (LSCM) and fluorescence activated cell sorter (FACS) were used.


RESULTS
Fluorescence microscopy revealed specific binding of FITC-BS to hybridoma cell. FACS analysis demonstrated that the binding was specific, saturable and reversibility. BS was used as target protein to screen its binding peptides from 12-mer random phage display peptide library. After four rounds of biopanning, 20 phage clones were randomly selected and identified. ELISA and competitive inhibition test results indicated that 2 phage clones were identified as positive clones. The amino acid sequences analysis shown that the sequences were ACTKHLCLLQPL or MSCNDTLCLLPN, which sharing a conservative sequence LCLL. In vitro experiments suggested that the two binding peptides can inhibit BS specific binding to hybridoma cell.


CONCLUSIONS
These results confirmed that existence of BS receptor in the membrane of hybridoma cell, which involved in hybridoma cell secreting monoclonal antibody signal pathway.
Wang
Chen
C
Lab of Veterinary Microbiology, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China. wangchen2001@126.com
Zhao
Zhanqin
Z
Niu
Mingfu
M
Zhang
Chunjie
C
Cheng
Xiangchao
X
Chen
Puyan
P
D000595:Amino Acid Sequence; D015870:Gene Expression; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D019151:Peptide Library; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*
D009842:Oligopeptides; D019151:Peptide Library; D011956:Receptors, Cell Surface; C016281:bursopoietin