人心肌型脂肪酸结合蛋白单克隆抗体的制备及定量检测试剂盒的研制

Preparation of monoclonal antibody against H-FABP and development of a quantitative ELISA kit in detection of H-FABP

目的：利用抗心肌型脂肪酸结合蛋白单抗，研制定量检测心肌型脂肪酸结合蛋白（ H-FABP ）的ELISA试剂盒。方法使用基因重组H-FABP免疫小鼠，以杂交瘤技术制备特异性抗H-FABP单抗，用这些单抗研制定量检测H-FABP的ELISA 试剂盒。结果筛选获得2株稳定分泌抗H-FABP单抗的杂交瘤细胞株，研制了定量检测H-FABP的ELISA试剂盒，灵敏度达到0．2 ng/mL，线性范围0．4～25 ng/mL，r2＝0．9967，回收率在97．2％～104．5％，精密度的变异系数（CV）≤6．72％；应用此试剂盒检测健康人血浆H-FABP，含量为1．87～8．50 ng/mL。结论所研制的ELISA试剂盒有较好的灵敏度及特异性，可用于人血浆中H-FABP含量的检测。

Objective To prepare monoclonal antibody against human heart-type fatty acid binding protein （ H-FABP ） and to establish a quantitative ELISA Kit in detection of H-FABP .Methods It was by immunization of mice with recombinant H-FABP ,and cell fusion was processed to establish hybridoma cell lines , the specific monoclonal antibodies against H-FABP were prepared .Results The quantitative ELISA in detection of H-FABP was developed by using these mono-clonal antibodies.The sensitivity of this assay reaches to 0.2 ng/mL, and the linear range is in 0.5-32 ng/mL with r2 val-ue of 0.996 7 and recovery rates are in 97.2%-104.5%,the CV of precision is less than 6.72%.It can measure H-FABP level in healthy human plasma in concentration 1.87-8.50 ng /mL.Conclusion The developed ELISA kit has better sensitivity and specificity , which could be used in detection of H-FABP in healthy human plasma .