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目的 :建立灵敏、特异和稳定的人可溶性CD4 0配体 (sCD4 0L)酶联检测试剂盒。方法 :采用鼠抗人CD4 0L单克隆抗体 1B1为包被抗体 ,识别人CD4 0L不同位点的单克隆抗体 4F1经琥珀酰羟基生物素 (BNHS)标记后为检测抗体 ,建立双抗体夹心的人sCD4 0L酶联检测方法。在此基础上 ,测定 2 0 0例健康供血员血清和血浆中sCD4 0L的含量。结果 :成功研制人sCD4 0L酶联检测试剂盒 ,灵敏度为 0 5ng ml。该试剂盒 4℃放置 3个月 ,离散度 (CV) <± 6 9% ,回收率为 94 7%～ 10 4 8% ,提示该检测系统具有良好的灵敏度、稳定性和准确性 ,与国外商品化试剂盒的分析结果相似。应用该试剂盒测得的健康供血员血清和血浆sCD4 0L含量分别为 9 5 6± 4 71、2 98± 1 13ng ml。结论 :研制成灵敏、特异和稳定的人sCD4 0L酶标检测试剂盒 ,能够对血清和血浆中sCD4 0L进行准确定量分析 ,并首次证实血清和血浆中sCD4 0L水平的差异性
Objective: To establish a sensitive, specific and stable human soluble CD4 0 ligand (sCD40L) enzyme linked detection kit. METHODS: Murine anti-human CD4OL monoclonal antibody 1B1 was used as the coating antibody to identify human CD4L monoclonal antibody 4F1 labeled with succinylhydroxy biotin (BNHS) as the detection antibody to establish a double antibody sandwich sCD4 0L enzyme-linked detection method. On this basis, the content of sCD40L in serum and plasma of 200 healthy blood donors was determined. Results: The human sCD40L ELISA kit was successfully developed with a sensitivity of 0.5 ng ml. The kit was placed at 4 ° C for 3 months with a CV of <± 6 9% and a recovery of 94.7% -104.8%, indicating that the system has good sensitivity, stability and accuracy. Analysis of commercial kits was similar. The sCD40L content of healthy blood donors measured by this kit was 956 ± 4 71,2 98 ± 1 13ng ml, respectively. Conclusion: A sensitive, specific and stable human sCD40L ELISA kit was developed to accurately and quantitatively determine sCD40L in serum and plasma, and it was the first time to confirm the difference of sCD40L levels in serum and plasma