CAJ | 학술논문

目的建立双抗体夹心ELISA法定量检测重组人干扰素α1b的方法.方法筛选具有不同抗原结合位点的抗重组人干扰素α1b单克隆抗体,分别作为包被抗体和辣根过氧化酶标记抗体,建立双抗体夹心ELISA法定量检测不同批次重组人干扰素α1b含量,评价该法的检出限、精确度、重复性、特异性.结果所建立的ELISA最低检出限为10 ng/ml,检测线性范围10～100 ng/ml,R2 =0.992,测定值与实际值偏差＞5％,板间变异系数均小于10％.结论该方法灵敏度高,特异性强,准确性和重复性好,可用于重组人干扰素α1b成品的定量检测.
목적 건립쌍항체협심ELISA법정량검측중조인간우소α1b적방법.방법 사선구유불동항원결합위점적항중조인간우소α1b단극륭항체,분별작위포피항체화랄근과양화매표기항체,건립쌍항체협심ELISA법정량검측불동비차중조인간우소α1b함량,평개해법적검출한、정학도、중복성、특이성.결과 소건립적ELISA최저검출한위10 ng/ml,검측선성범위10～100 ng/ml,R2 =0.992,측정치여실제치편차＞5％,판간변이계수균소우10％.결론 해방법령민도고,특이성강,준학성화중복성호,가용우중조인간우소α1b성품적정량검측.
Objective To develop a double antibody sandwich ELISA assay for quantitative determination of recombinant human interferon α1b.Methods Mouse monoclonal antibodies with different binding site on rIFN-α1b were screened to select optimized candidates as coating and HRP-labeled index antibodies respectively.And a double antibodies sandwich ELISA was assembled; the reliable lower detection limit,specificity,accuracy and reproducibility were evaluated and validated.Results The quantitative sandwich ELISA had a reliable lower detection limit of 10 ng/ml,with a liner detection range 10-100 ng/ml (R2 =0.992),variation coefficient inter-plates is less than 10％.Conclusion The developed sandwich ELISA was a sensitive and specific,accuracy and reproducibility method for quantitative determination of recombinant human interferon αt1b in final product.