The mimic epitope of N-methyl-D-aspartate receptor 2B subunit screened from peptide library displayed on phage / 中华麻醉学杂志

ABSTRACT

Objective To determine the mimic epitope of N-methyl-D-aspartate receptor 2B subunit (NR2B). Methods The monoclonal antibody interacting with NR2B was used as target protein to screen the binding peptide from a 12-mer Ph. D random peptide library. After three rounds of affinity screening, twelve specific clones were selected randomly and identified by sandwich ELISA. The peptide sequences were analyzed by DNA sequencing. The clones containing a common sequence were named positive clone. The competitive inhibition of the native antigen bound to monoclonal antibody against the NR2B by the positive clone was assayed by cell ELISA. Results After 3 rounds of screening the phages specifically bound with mAb NR2B were selected and amplified. Nine of the 12 clones displayed a common aminoacid sequence SHPPVMPWPTST. The inhibitory assay showed that the mimic epitope peptides displayed on the phage surface could effectively inhibit the combination of the monoclonal antibody with native antigen. The inhibitory rate of mimic epitope was (45?3)% .Conclusion The mimic epitope of NR2B was screened successfully from the 12-mer Ph D random peptide library. This peptide mimics an epitope of the native NR2B.