Comparing ELISA and surface plasmon resonance for assessing clinical immunogenicity of panitumumab
Comparing ELISA and surface plasmon resonance for assessing clinical immunogenicity of panitumumab
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DOI：
10.4049/jimmunol.178.11.7467
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发表时间：
2007-06-01
影响因子：
4.4
通讯作者：
Mullenix, Michael C.
中科院分区：
文献类型：
--
作者：
Lofgren, James A.;Dhandapani, Sripriya;Mullenix, Michael C.
Panitumab是一种完全人源性的抗表皮生长因子受体单抗，被批准用于结直肠癌患者，对其免疫原性的评估导致了两种独立的检测Panitumab抗体的免疫分析方法的发展。第一次免疫检测采用桥接ELISA法，可检测10 ng/ml阳性对照抗Panitumumab抗体。该酶联免疫吸附试验采用了酸解离步骤以减少药物干扰，并可耐受类似摩尔过量100倍药物的存在。在8项临床试验中，ELISA检测到612名受试者中有2名(0.3%)出现了抗体反应。在其中一名ELISA阳性受试者中，用表皮生长因子受体磷酸化生物法检测中和抗体。第二种免疫分析采用Biacore生物传感器免疫分析形式，能够在耐受等摩尔量药物存在的情况下检测1微克/毫升阳性对照抗体。虽然在试验中对竞争药物的敏感性和耐受性较低，但Biacore试验在604名受试者中有25人(4.1%)检测到发展中的抗体反应。此外，Biacore试验确定了8名形成中和抗体的受试者。亲和力为1.1×10~(-6)~8.4×10~(-10)M的鼠单抗用于鉴定这两种方法。该方法对高亲和力单抗的检测更为敏感，当药物与单抗的摩尔比较高时，可检出高亲和力单抗。Biaccore法对低亲和力单抗的检测更敏感，当药物与单抗的摩尔比较高时，可检测到低亲和力单抗。
Evaluation of the immunogenicity of panitumumab, a fully human anti-epidermal growth factor receptor mAb approved for use in colorectal cancer patients, led to the development of two separate immunoassays for the detection of anti-panitumumab Abs. The first immunoassay used a bridging ELISA capable of detecting 10 ng/ml positive control anti-panitumumab Ab. The ELISA incorporated an acid dissociation step to reduce drug interference and tolerated the presence of similar to 100-fold molar excess of drug. During eight clinical trials, the ELISA detected developing Ab responses in 2 of 612 (0.3%) subjects. In one of the ELISA positive subjects, neutralizing Abs were detected using an epidermal growth factor receptor phosphorylation bioassay. The second immunoassay used a Biacore biosensor immunoassay format capable of detecting 1 mu g/ml positive control Ab while tolerating the presence of equal molar amounts of drug. Although less sensitive and less tolerant to competing drug in the assay, the Biacore assay detected developing Ab responses in 25 of the 604 (4.1%) subjects. Additionally, the Biacore assay identified eight subjects who developed neutralizing Abs. Mouse mAbs with affinities ranging from 1.1 X 10(-6) to 8.4 X 10(-10) M were used to characterize both assay types. The ELISA was more sensitive for the detection of higher affinity mAbs and detected high-affinity mAbs in the presence of higher molar ratio of drug to mAb. The Biacore assay was more sensitive for detection of lower affinity mAbs and detected low affinity Abs in the presence of higher molar ratios of drug to mAb.