摘要
目的制备兔抗Cap43多克隆抗体。方法以30个氨基酸组成的多肽为半抗原与钥孔血蓝蛋白(mcKLH)通过碳化二亚胺(EDC)法化学偶联成完全抗原(多肽-mcKLH)免疫家兔;制备兔抗Cap4443多克隆抗体;斑点ELISA法检测抗体效价,经辛酸-硫酸铵法初步纯化抗体后,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)电泳考马斯亮蓝检测其纯化后的纯度。结果经免疫11周得到兔抗Cap43多克隆体,抗体效价为1:500。结论采用EDC化学偶联半抗原和大分子载体蛋白mcKLH,使其成为同时具有免疫原性与免疫反应性的完全抗原,用其免疫家兔获得多克隆抗体。
Objective To obtain an anti - Cap43 polyclonal antibody. Methods Peptide with 30 amino acids as hapten was coupled with mcKLH using carbodiimide （EDC） coupling chemical technique to form a coupling agent （peptidemcKLH）, then rabbits were immunized;To prepare anti - Cap43 polyclonal antibody. Valence of antibody was detected by dot - ELISA. The polyclonal antiserum was purified by caprylic acid - ammonium sulfate precipitation, then, the purity of antigen was detected by SDS- PAGE. Results Rabbit polyclonal antiserum against human Cap43 protein was obtained after 11 weeks and the titer of the antiserum obtained in this experiment was up to 1 ： 500. Conclusion Coupling hapten with giant molecular vehicle mcKLH by EDC can make it complete antigen that posessed immunogenicity and immunoreactivity. Immunizing rabbits with the hapten - mcKLH was used to obtain the antibody against Cap43.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2007年第3期289-291,共3页
Chinese Journal of Public Health
基金
国家自然科学基金(30471474)
