关键词: 双峰驼, IgG亚型, 分离纯化, 多克隆抗体

Abstract: IgG subclasses of Bactrian camel is consist of IgG1, IgG2 and IgG3. Both IgG2 and IgG3 are the heavy chain antibodies (HCAbs), with structures that are obviously different from IgG1’s. To obtain IgG1, IgG2 and IgG3 and analyze their antigen specificities, IgG subclasses were isolated and purified from the Bactrian camel serum using Protein A and Protein G affinity chromatography columns, and were identified by polyacrylamide gel electrophoresis (SDS-PAGE). Then polyclonal antibodies to them were prepared respectively and their titers were determined by enzyme-linked immunosorbent assay (ELISA). Finally, their specificities were evaluated by Western blot, and the antigen-specificity of IgG subclasses of Bactrian camel were further analyzed. The results showed that the IgG1, IgG2 and IgG3 of Bactrian Camel are obtained successfully by the Protein A and Protein G affinity chromatography columns, the titers of the polyclonal antibodies of anti-IgG1, IgG2, and IgG3 were all over 1:10000, and every polyclonal antibody had obvious cross-reactivity with IgG1, IgG2 and IgG3, but the polyclonal antibody specificity of anti-IgG1 was lower than those of anti-IgG2 and anti-IgG3. The results suggested that the immunogenicity of IgG1, IgG2 and IgG3 of Bactrian camel were all good and their specificities were all similar although their structures were obviously different.

Key words: IgG subclasses, Bactrian camel, Isolation and purification, Polyclonal antibody