The Preparation Of PAG1 Monoclonal Antibodies

Pregnancy-associated glycoproteins(PAG)are macromolecules produced by placental tissue and released into the maternal circulation where they allow pregnancy diagnosis and Follow-up.Recent studies focuses on the indirect pregnancy detection methods that unharmful,high accuracy and strong specificity,such as pregnancy associated glycoproteinl(PAG1)detection in blood.In order to develop products just like human early pregnancy test strip to obtain an accurate,rapid method of PAG1 content detection in serum,first of all we need a kind of strong specificity PAG1 monoclonal antibody,and lay the foundation for development of PAG 1 colloidal gold test strip.The contents and results of this experiment are as follows:(1)The PAG1 recombinant protein prokaryotic expression system was successfully constructed according to the PAG1 amino acid sequence of NCB1,and the recombinant PAG1 antigen was expressed in a large number of recombinant strains.Results:the position of PAG1 recombinant protein expressed in E.coli was correct;(2)PAG1 polyclonal antibodies were prepared by immunization of New Zealand white rabbits with PAG1 antigen and PAG1 recombinant antigens.Results:the titer of ELISA result shows polyclonal antibody of PAG 1 was obtained;(3)Balb/c SPF female mice were immunized with PAG1 antigen and PAG1 recombinant antigens,The spleen cells from immunized mice were fused with SP2/0 cells using PEG technique.And then screen and identify the monoclonal antibodies.Results:39 monoclonal cell lines of PAG 1 were selected,and the monoclonal antibody of IgG was successfully obtained.Conclusion:We successfully expressed recombinant PAG1 protein through the prokaryotic expression system,successfully prepared the polyclonal antibodies and monoclonal cell strains against different epitopes of PAG1 and lay the foundation for the following research.