Expression and preparation of polyclonal antibodies against human apolipoprotein A5 and tissue distribution / 中国实用内科杂志

ABSTRACT

Objective To prepare the recombination protein and polyclonal antibodies against human apolipoprotein A5(ApoA5)and detect the expression of ApoA5 in human tissues with the antibody,then further study the fuction and serum level of ApoA5.Methods With PCR and gene recombinated techniques,the prokaryotic ApoA5 expression vector was constructed.The His-ApoA5 fusion protein was expressed in E.ColiM15,and the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody.The antibody was tested for their specificity with ELISA and Western-blot,then it was used to detect the tissue distribution of ApoA5.Results It was found that there was a specific brand at 40 ku of the recombination ApoA5(rApoA5);the polyclonal antibody had high immunoreactivity and specificity.ApoA5 was expressed in human sera and hepatic tissue,not in other tissues(heart,vessel,spleen,intestine and lung),regardless of the lipid level.Conclusion The pQE30-ApoA5 is constructed successfully,the rApoA5 can be expressed in E.Coli M15;ApoA5 antibody has highly specifity.It can be used in clinical test.There is ApoA5 in human serum,and ApoA5 is expressed in liver which may be significant in lipid metabolism.