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 |
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引用本文: | 张加利,蔡芳,高强,宋莉莉,于丹,桑建利.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[J].粉末涂料与涂装,2008,21(11). |
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作者姓名: | 张加利 蔡芳 高强 宋莉莉 于丹 桑建利 |
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作者单位: | 北京师范大学生命科学学院,北京科兴生物制品有限公司质量管理部 |
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摘 要: | 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。
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关 键 词: | 牛血清白蛋白 单克隆抗体 双抗体夹心ELISA法 |
Screening of Monoclonal Antibody against Bovine Serum Albumin and Development of Double Antibody Sandwich ELISA for BSA |
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Abstract: | 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. |
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Keywords: | Bovine serum albumin Monoclonal antibody Double antibody sandwich ELISA |
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