Murray G J, Howard K D, Richards S M, Barton N W, Brady R O
Developmental and Metabolic Neurology Branch National Institutes of Health, Bethesda, MD 20892.
J Immunol Methods. 1991 Mar 1;137(1):113-20. doi: 10.1016/0022-1759(91)90400-a.
An amplified ELISA has been employed for monitoring the safety of repeated intravenous infusions of modified human placental glucocerebrosidase. The enzyme infusions consisted of biweekly injections of macrophage targeted glucocerebrosidase over a 6 month duration. Serum samples collected throughout the study were assayed by use of an ELISA using alkaline phosphatase coupled to alcohol dehydrogenase for amplification. Using this protocol, 0.2-5 ng affinity purified immunoglobulin specific for glucocerebrosidase can be detected. Occasional false positives necessitate multiple repeat assays over time to accurately assess immunogenic response. Blinded ELISAs were performed on sera from both infused patients with Gaucher's disease and uninfused control patients and compared with apparent immunoglobulin concentration in 54 normal control sera. Although several samples showed apparently elevated immunoglobulin levels, repeat analyses failed to demonstrate high levels reproducibly. Furthermore, these sera were unable to neutralize enzyme or to precipitate radiolabelled enzyme, confirming the absence of antibody. Problems with high sensitivity ELISA formats are discussed.
一种放大酶联免疫吸附测定法（ELISA）已被用于监测经修饰的人胎盘葡萄糖脑苷脂酶重复静脉输注的安全性。酶输注包括在6个月的时间内每两周注射一次靶向巨噬细胞的葡萄糖脑苷脂酶。在整个研究过程中收集的血清样本通过使用与乙醇脱氢酶偶联的碱性磷酸酶进行放大的ELISA进行检测。使用该方案，可以检测到0.2 - 5纳克对葡萄糖脑苷脂酶具有特异性的亲和纯化免疫球蛋白。偶尔出现的假阳性需要随着时间进行多次重复检测，以准确评估免疫原性反应。对患有戈谢病的输注患者和未输注的对照患者的血清进行了盲法ELISA检测，并与54份正常对照血清中的表观免疫球蛋白浓度进行了比较。尽管有几个样本显示表观免疫球蛋白水平明显升高，但重复分析未能再现性地证明高水平。此外，这些血清无法中和酶或沉淀放射性标记的酶，证实不存在抗体。讨论了高灵敏度ELISA检测形式存在的问题。
