牛血清白蛋白单克隆抗体的筛选与双抗体夹心ELISA检测方法的建立

Screening of Monoclonal Antibody against Bovine Serum Albumin and Development of Double Antibody Sandwich ELISA for BSA

目的筛选高效抗牛血清白蛋白(BSA)单克隆抗体细胞株,并建立BSA抗原检测的双抗体夹心ELISA法。方法分别采用辛酸-硫酸铵法和葡萄球菌A蛋白亲和层析法纯化单抗,并进行抗体类型、腹水效价、特异性和相对亲和力测定;应用纯化的单抗建立BSA双抗体夹心ELISA检测法,并进行初步应用。结果筛选出4株可稳定分泌抗BSA单抗的杂交瘤细胞株,抗体类型为IgG,抗体滴度均可达10-6,特异性良好,相对亲和力较高。建立的双抗体夹心ELISA法线性范围为1.25～20ng/ml,R2>0.98,灵敏度为1.25ng/ml。结论已筛选出高效抗BSA的单抗,并建立了BSA双抗体夹心ELISA检测方法。

Objective To screen the monoclonal antibody （McAb） against bovine serum albumin （BSA） and develop a double antibody sandwich ELISA for BSA. Methods The McAb was purified by eaprylic acid-ammonium sulphate precipitation and staphylococcal protein A affinity chromatography and analyzed for type, titer in ascites, specificity and relative affinity. A double antibody sandwich ELISA was developed with the purified McAb and applied primarily. Results Four hybridoma cell strains stably secreting MeAb against BSA was screened. The secreted McAb with a titer of 10^-6 was IgG which showed good specificity and high relative affinity. The sensitivity of developed double antibody ELISA was 1. 25 ng/ml, and the linear range and R^2 value of detection result by the method were 1.25 - 20 ng/ml and more than 0. 98 respectively. Conclusion The McAb against BSA was suceessfully screened, and a double antibody sandwich ELISA for BSA was developed.