| The study choose grass carp as the experimental object. â‘ Based on the deduced amino-acid sequence, gcGH were compared with known ccGH in this experiment. eleven peptides representing sequences from gcGH and an extra cystine added to its N-terminal part were chosen to synthesis based on gcGH and ccGH having minimal sequence identity, maximal hydropathy of gcGH, and maximal probability of corresponding to strongly immunogenic region of gcGH, design specific antibodies useful for detection of grass carp growth hormone;â‘¡Twelve peptides were corresponding to solid-phase methodology on an ABI 430A peptide synthesizer employing user-devised cycles the crude peptides were purified to >96% by reversed-phase HPLC with a gradient of acetonitrile in struture and purity were confirmed in all cases by electrospray ionization-mass spectrometry; â‘¢SC-12 is a kind of hapten that must conjugate to carrier protein KLH with MBS methods;â‘£gcGH polycolonal antibody system is established; highly titer and specifc antiserum is prepared;the results as follows: Western blot and cross reaction of gcGH and ccGH with gcGH antiserum of 1:10000 dilution confirmed that gcGH protein could specifically react with antibody against gcGH.A method that determines the immune quantification study of GH expressed has been successfully established with the antigen peptide in this study. And then by this method through the specific combination between antigen and antibody, it has been further confirmed that the anti-serum has the specific reaction to the active grass carp growth hormone. With the ELISA and Western blot technique, we carried out the immune quantification study of gcGH that will be the basis to develop a method to determine the GH and expressed protein. The result of this study can provide the scientific data to develop a rapid, accurate and sensitive detection method of grass carp, and is important to the study of molecular biology of fish growth hormone and GH gene transfer breeding.
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