摘要
目的:SARS 冠状病毒(SARS-CoV)核衣壳蛋白 N(nucleocapsid protein)单克隆抗体(monoclonal antibody,McAb)的制备及鉴定。方法:用基因重组的 SARS-CoV N 蛋白免疫 BALB/c 小鼠制备 McAb,并采用间接 ELISA、免疫印迹法进行筛选和鉴定。结果:筛选出2株抗 SARS-CoV N 蛋白 McAb 杂交瘤细胞株,间接 ELISA 证实这组单克隆抗体仅与 SARS-CoV 产生特异性反应,而与其他病原体无交叉反应,IgG 亚类鉴定1株为 IgG1,另1株为 IgG2b。2株抗体亲和常数分别为4.14×10^(-9)和3.19×10^(-9)。结论:获得特异性针对 SARS-CoV 的单克隆抗体,为 SARS-CoV 早期诊断试剂的研制奠定了基础。
Objective：To develop a method for preparing and identifying monoclonal antibodies （McAb） against SARS - associated coronavirus （ SARS - CoV） nucleocapsid （N） protein. Methods ：BALB/c mice were injected with the recombinant N protein of SARS - CoV into the hypophloeodal for the immunization, and the splenic cells were isolated 15 d later for McAb -producing hybridomas,from which the McAbs against the N protein of SARS -CoV were screened. The identification of the McAb against the N protein of SARS - CoV was performed using indirect enzyme - linked immunosorbent assay （ELISA）, and Western immunoblotting. Results：Two strains of hybridomas were obtained that produced the McAb specific to the N protein without detectable cross - reactivity with other pathogens. One was identified as the immunoglobulin G1 （IgG1） isotype, the other IgG2a,with affinity constants （Ka） of two strains being 4. 14 ×10^-9 and 3. 19×10^-9 respectively. Conclusion：This is the first report on the preparation of McAb that is specific to the SARS - CoV, and the high - specificity and high - affinity McAb produced by the two strains of hybridomas provide a basis for further researches on the pathogenesis and early diagnosis of SARS.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2007年第8期1292-1294,共3页
Chinese Journal of Pharmaceutical Analysis
关键词
严重急性呼吸综合征
冠状病毒
核衣壳蛋白
单克隆抗体
Sever Acute Respiratory Syndrome
Coronavirus
Nucleocapsid protein
Monoclonal antibody
