Labus J M, Petersen B H
Lilly Laboratories for Clinical Research, Indianapolis, Indiana 46285.
Cytometry. 1992;13(3):275-81. doi: 10.1002/cyto.990130309.
Clinical investigations utilizing murine monoclonal antibodies require techniques for the detection of the human anti-mouse antibody (HAMA) response in patient serum. We report here a flow cytometric assay for the quantitation of HAMA. Commercially available beads conjugated with goat anti-mouse antibody provide a solid phase matrix for a triple bridge immunoassay. The measurement of fluorescein labeled antibodies by flow cytometry allows accurate quantitation of the HAMA. The assay will detect antibody levels of approximately 1.0 ng/ml. Antibody recovery in serum samples with known amounts of antibody added was greater than 90% at levels greater than or equal to 10 micrograms/ml. Serum samples obtained from 41 patients prior to and after single or multidose infusions of KS1/4-Desacetyl-vinblastine were analyzed. These results were compared with HAMA titers previously determined by ELISA. With few exceptions, patients with low titers as determined by ELISA demonstrated low HAMA potencies by flow cytometry and those with highest titers the highest potencies. Patients with no detectable HAMA by ELISA were also negative by flow analysis. The results of our studies demonstrate that HAMA levels can be accurately and quantitatively determined by flow cytometry.
利用鼠单克隆抗体的临床研究需要检测患者血清中人类抗鼠抗体（HAMA）反应的技术。我们在此报告一种用于定量HAMA的流式细胞术检测方法。与山羊抗鼠抗体偶联的市售微珠为双桥免疫测定提供了固相基质。通过流式细胞术测量荧光素标记的抗体可实现对HAMA的准确定量。该检测方法可检测到约1.0 ng/ml的抗体水平。在添加已知量抗体的血清样本中，当抗体水平大于或等于10微克/毫升时，抗体回收率大于90%。分析了41例患者在单次或多剂量输注KS1/4-去乙酰长春碱之前和之后采集的血清样本。将这些结果与先前通过ELISA测定的HAMA滴度进行比较。除少数例外情况外，ELISA测定为低滴度的患者通过流式细胞术显示出低HAMA效价，而滴度最高的患者效价也最高。ELISA检测不到HAMA的患者通过流式分析也为阴性。我们的研究结果表明，通过流式细胞术可以准确、定量地测定HAMA水平。
