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

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。

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

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 caprylic acid-ammonium sulphate precipitation and staphylococcal protein A affinity chromatography and analyzed for type,titer in ascites,specificity and relative affinity.A double anti-body sandwich ELISA was developed with the purified McAb and applied primarily.Results Four hybridoma cell strains stably se-creting McAb against BSA was screened.The secreted McAb with a titer of 10-6 was IgG which showed good specificity and high rel-ative affinity.The sensitivity of developed double antibody ELISA was 1.25 ng / ml,and the linear range and R2 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 successfully screened,and a double antibody sandwich ELISA for BSA was developed.