Expression of bursin receptor on membranes of hybridoma cell and Identification of binding peptide of bursin from 12-merrandom phage display peptide library

Abstract:

Abstract: [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. [Conclusion] These results confirmed that existence of BS receptor in the membrane of hybridoma cell, which involved in hybridoma cell secreting monoclonal antibody signal pathway.