摘要
目的：建立一种灵敏、特异、快速的ELISA方法,用于检测食蟹猴血清中重组抗CD52单克隆抗体的含量。方法：采用双抗夹心ELISA法对重组抗CD52单克隆抗体进行定量。以猴血清吸附的羊抗人Ig G作为包被抗体,稀释的猴血清吸附的羊抗人Ig G-HRP（二抗）作为检测抗体,加入底物显色剂后在酶标仪上读取D450nm值。结果：建立并确证了检测重组抗CD52单克隆抗体的ELISA方法,方法的线性范围为7.81~500 ng/m L,定量下限为7.81 ng/m L,板内及板间精密度和准确度均在±15%以内,室温、冻融、稀释效应稳定性良好。结论：方法学验证表明ELISA法测定食蟹猴血清中重组抗CD52单克隆抗体浓度的特异性、精密度和准确度均满足新生物制药临床前药代动力学研究指导原则要求,可用于重组抗CD52单克隆抗体的检测。
Objective： To establish a specific, sensitive and rapid enzyme linked immunosorbent assay（ELISA） method for detection of rh-anti-CD52ximAb in rhesus monkey serum. Methods： This study employed the quantita- tive sandwich enzyme immunoassay technique. The assay method utilized sheep anti-human IgG that had been ad- sorbed against cynomolgus monkey serum for capture as well as detection of rh-anti-CD52ximAb. Following that, color was developed by the substrate solution and the reaction was stopped by stop solution. Then the plate was analyzed with a microplate reader at a wavelength of 450 nm. Results： An ELISA assay was developed with a wide dynamic range of concentrations from 7.81 to 500 ng/mL with a lowest quantification of 7.81 ng/mL, both ac- curacy of the intra- and inter-assay were less than 15%. Meanwhile, well stability of rh-anti-CD52ximAb was shown by evaluation on different storage conditions and dilution with serum. Conclusion： This method is highly sensitive, accurate, specific, and reproducibility, which was proven to be a feasible quantitative method for the de- tection of rh-anti-CD52ximAb.
出处
《生物技术通讯》
CAS
2015年第5期667-671,共5页
Letters in Biotechnology
