摘要
采用杂交瘤法制备单克隆抗体,并用辛酸-硫酸铵法纯化单抗,通过ELISA方法和Western blotting测定抗体的效价与特异性,并进行抗体类型、相对亲和力测定;应用纯化的单抗建立hGH双抗体夹心ELISA检测方法。筛选出两株可以稳定分泌抗hGH单抗的杂交瘤细胞株,分别命名为3E11、2G9,抗体类型均为IgG1,抗体滴度均可达10-10,特异性好,相对亲和力高,以筛选到的两株单抗建立的双抗夹心ELISA法线性范围为0.09～1.5625ng/mL,R2>0.9,灵敏度为0.09ng/mL。筛选出高效抗hGH的单抗,并建立了hGH双抗体夹心ELISA检测方法。
To screen the monoclonal antibody（McAb） against recombinant human growth hormone（rhGH） and develop a double antibody sandwich ELISA for hGH,the McAb was prepared by using hybridoma technique and purified by caprylic acid-ammonium sulphate precipitation.The McAB was analyzed for type,titer in ascites,relative affinity and specificity.By using the pair of McAbs,a double antibody sandwich ELISA method was established.Two hybridoma cell strains stably secreting McAb against hGH were screened.The secreted McAbs with a titer of 10-6 and 10-7 was IgG1 with good specificity.The linear range and R2 value of detection result were 0.09ng/mL and more than 0.99 respectively,the sensitivity of developed double antibody sandwich ELISA was 0.09ng/mL.
出处
《生物学杂志》
CAS
CSCD
2011年第5期94-97,共4页
Journal of Biology
关键词
人生长激素
单克隆抗体
双抗体夹心ELISA法
human growth hormone
monoclonal antibody（McAb）
double antibody sandwich ELISA
