Screening Of The Antigenic Epitope Of Pgh By Using Phage Displayed Random Peptide Library

Porcine grown hormone is a peptide hormones secreted by the pituitary,and functions related to growth and reproduction.a large number of studies have show that porcine growth hormone can significantly improve grownth rate,leand meat and feed in the process of growth and development,significant impact on porcine production,breeding and deployment of feed. Our study is research for epitope of porcine growth hormone. The main results of this study are as follows:1.Using the hhybridoma that preparation by our laboratory, injection into BALB/c mice which production after immunization by liquid paraffin a week.uesed acid-crude saturated ammonium sulfate method to caprylicing mice ascites, and then purified by chromatography system protein,and finally test purity with SDS-polyacrylamide gel electrophoresis, indirect ELISA detection the OD490 values, titer of hybridoma culture supernatant 1:1600, the obtained ascites titer is 1:1.6 x 1042. A phage-displayed random 12-mer peptides library was screened four rounds with porcine growth hormone monoclonalantibody as target, coefficient of recovery of phage clone increased from 1.1×10-81.0×10-2, the targets phage clones have been effectively enriched. randomly picked 47 blue plaques to amplified then detected by indirect ELISA, found there were nine plaque clones has strong specificity with the monoclonal antibody,take ssDNA from nine positive clones for Sequencing and alignment analysis, the result show that seven positive clones of exogenous peptide sequence completely consistent with the amino acid sequences were compared and obtained homologous sequences NxxxTSx. These mimic epitope with a certain reference value to the porcine growth hormone epitopes. Lay the foundation for further analysis the structure and function of porcine growth hormone.