动物造模 z-bio 2022-05-25 245

　　OBJECTIVE: To purify IgG antibody from marmoset serum, prepare rabbit anti-marmoset antiserum, and purify IgG from antiserum, HRP (horseradish peroxidase)-labeled rabbit anti-marmoset IgG.

　　Methods: HiTrap™ Protein G affinity chromatography was used to purify marmoset and rabbit anti-marmoset serum IgG. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used for purity identification. Assay) to determine the prepared rabbit anti-marmoset antiserum titer, improve the "simple sodium periodate labeling method" to prepare rabbit anti-marmoset IgG-HRP-labeled antibody, enzyme-linked immunosorbent assay (ELISA) and Western blotting ( Western blotting) to determine the working concentration and specificity of the rabbit anti-marmoset IgG-HRP-labeled antibody.

　　RESULTS: The purity of IgG purified from marmoset and rabbit anti-marmoset sera was greater than 95% and 97%, respectively; the titer of rabbit anti-marmoset IgG antiserum was 1∶64. ELISA and Western blotting identified the reference working concentration of rabbit anti-marmoset IgG-HRP enzyme-labeled antibody at 1:256 000 and 1:15 000, with obvious specificity.

　　Conclusion: The preparation and preliminary identification of marmoset IgG-HRP-labeled antibodies, including the specificity and concentration of ELISA and Western blotting, reserve resources for the immunological detection system and molecular immunological detection system of marmoset pathogens.