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目的 :探讨用重组可溶性细胞间黏附分子 -1(sICAM -1)I～Ⅲ功能区蛋白免疫家兔制备抗体 ,及用其建立人血清sICAM -1放免方法 (RIA)。方法 :对获得的抗体和建立的RIA方法学进行鉴定。测定214例正常人和38例初诊Grave’s病 (GD)患者的sICAM -1。结果 :抗体的亲和常数为1 056×108L/mol ,最终稀释度为1∶14000。灵敏度为1 4μg/L ;非特异结合<5.0 % ;批内变异<5 % ,批间变异<8 % ;样品平均回收率为102 52 % ;平行实验的稀释曲线与标准曲线基本平行 ,参考值范围 (x±2s)为 (175 15±66 88) μg/L ,各年龄、性别组间差别无统计学意义(P>0.05) ,初诊GD患者血清sICAM -1显著高于对照组(P<0.01)。结论 :成功制备出抗体 ,用其建立了人血清sICAM -1RIA ,具有临床应用前景
OBJECTIVE: To investigate the immunization of rabbits with recombinant soluble intercellular adhesion molecule-1 (sICAM-1) I ~ III protein and preparation of antibodies against human sICAM-1 by radioimmunoassay (RIA). Methods: The acquired antibodies and established RIA methodology were identified. SICAM-1 was measured in 214 normal subjects and 38 newly diagnosed Grave’s disease (GD) patients. Results: The affinity constant of the antibody was 1 056 × 108 L / mol and the final dilution was 1:14 000. The sensitivity was 14μg / L; non-specific binding was less than 5.0%; intra-assay variation was less than 5%, intra-assay variation was less than 8%; the average recovery was 102.52%. The dilution curve of the parallel experiment was basically parallel to the standard curve, The range of (± 2s) was (175 15 ± 66 88) μg / L, and no significant difference was found between the age and sex groups (P> 0.05). The sICAM-1 of newly diagnosed GD patients was significantly higher than that of the control group (P < 0.01). Conclusion: The antibody was successfully prepared and used to establish human serum sICAM-1RIA, which has the clinical application prospect