人可溶性B7-2分子的定量检测及应用

选取识别不同抗原位点的鼠抗人B7-2单抗1F9和6C8分别作为包被和检测抗体,用生物素(Biotin)标记检测抗体,建立双单抗夹心的sB7-2检测方法。在对其特异性、稳定性和准确性进行分析的基础上,对20例系统性红斑狼疮(SLE)患者及30名健康献血者血清中sB7-2的含量进行了测定。建立的sB7-2检测方法的灵敏度为0.15 ng/ml,具有良好线性关系的检测范围0.31~20.00 ng/ml。单抗1F9包被的测定板,4℃放置30 d内,工作曲线中各测定点的变异系数<5.9%。SLE患者血清中sB7-2的含量为(2.207±0.517)ng/ml,与健康献血者的(1.275±0.263)ng/ml比较具有统计学的显著差异(p<0.01)。本研究中建立的人sB7-2检测方法,能够对血清中sB7-2进行定量分析,可为该分子的基础研究及临床相关疾病的辅助诊断与判断预后提供参数。

The quantitative determination of human soluble B7-2 molecule and its application

A method of quantitative determination of human soluble B7-2 molecule was established by using the mouse anti-human B7-2 monoclonal antibody 1F9 and 6C8 recognizing different antigenic epitopes on this molecule as the coating and detecting antibodies and the biotin-labeled monoclonal antibody in the double monoclonal antibody sandwich ELISA assay.The fusion protein B7-2/Fc was used as the standard antigen.On the basis of analysis in its specificity,stability and accuracy,the contents of soluble B2 molecule in sera of 20 cases with systemic lupus erythematosus(SLE) and 20 healthy volunteers were determined by using this method.It was found that this method was proved to be quite sensitive,specific and stable,with a detecting limit of 0.15 ng/ml and a better linear range of standard curve of 0.31～20.00 ng/ml.As determined by this method,no cross reaction could be detected with the monoclonal antibody against B7-1Fc.The concentrations of B72 in sera from SLE and healthy volunteers were(2.207 ± 0.517) ng/ml and(1.275 ± 0.263) ng/ml respectively,with significant statistical difference between these two groups(P<0.01).This method may provide further data of serological diagnosis and prognosis for the related diseases.