Biologics Clinical Pharmacology, Centocor Research and Development Inc., 145 King of Prussia Road, Radnor, PA 19087, USA.
J Pharm Biomed Anal. 2011 Jan 25;54(2):286-94. doi: 10.1016/j.jpba.2010.08.022. Epub 2010 Sep 24.
We assessed the utility of the FortéBio Octet(®) system for detection of anti-drug antibodies (ADAs) against an investigational therapeutic human IgG1 monoclonal antibody (mAb), CNTO X. To understand the relative merits of this technology, key performance requirements were compared with two popularly accepted ADA detection methods, a step-wise bridging ELISA and a Meso Scale Discovery (MSD) homogeneous (single step binding) bridging ECLIA. When used to detect 13 monoclonal ADAs of varying affinities and one polyclonal ADA, all three methods demonstrated their greatest apparent sensitivity to the polyclonal sample (1, 6, and 130 ng/mL, respectively for ECLIA, ELISA, and Octet). Sensitivity to monoclonal ADAs tended to vary in accordance with their affinities, however, the sensitivity of the Octet method varied much less between ADAs. As a result, the above ranking became reversed such that Octet was the most and ELISA least sensitive for detection of low-affinity ADAs. With regard to drug tolerance, the presence of CNTO X could lead to false-negative assay results, although each method was affected to a different degree, with the Octet method tolerating up to 10 times more drug than the ECLIA method, which in turn tolerated up to 10 times more than the ELISA. Finally, the ECLIA and Octet methods were applied to the bioanalysis of cynomolgus monkey sera from a pre-clinical multiple dose study of CNTO X. Octet indicated 3 positive animals developed ADA as early as day 15 of the dosing phase while drug was present at nearly 1mg/mL. ECLIA detected only one of these, and only in a day 57 recovery sample after drug had cleared from circulation. We conclude that the Octet is a promising platform for detection of lower affinity ADAs and is particularly suitable for ADA detection when drug persists at levels that negatively impact bridging immunoassays.
我们评估了 FortéBio Octet（®）系统用于检测针对一种研究性治疗性人 IgG1 单克隆抗体（mAb）CNTO X 的抗药物抗体（ADA）的效用。为了了解这项技术的相对优势，我们将关键性能要求与两种广受欢迎的 ADA 检测方法进行了比较，即逐步桥接 ELISA 和 Meso Scale Discovery（MSD）均相（单步结合）桥接 ECLIA。当用于检测 13 种具有不同亲和力的单克隆 ADA 和一种多克隆 ADA 时，所有三种方法都显示出对多克隆样品的最大敏感性（ECLIA、ELISA 和 Octet 分别为 1、6 和 130ng/mL）。对单克隆 ADA 的敏感性往往与其亲和力有关，但 Octet 方法在不同 ADA 之间的敏感性变化要小得多。因此，上述排名发生了逆转，即 Octet 对低亲和力 ADA 的检测最敏感，而 ELISA 最不敏感。关于药物耐受性，CNTO X 的存在可能导致假阴性检测结果，尽管每种方法的影响程度不同，Octet 方法耐受的药物比 ECLIA 方法多 10 倍，而 ECLIA 方法又比 ELISA 方法多耐受 10 倍。最后，我们将 ECLIA 和 Octet 方法应用于 CNTO X 临床前多次剂量研究的食蟹猴血清的生物分析中。Octet 表明，在给药阶段存在接近 1mg/mL 的药物时，早在第 15 天就有 3 只阳性动物产生了 ADA。ECLIA 仅检测到其中一种，并且仅在药物从循环中清除后的第 57 天恢复样品中检测到。我们得出结论，Octet 是一种很有前途的低亲和力 ADA 检测平台，尤其适用于药物持续存在且对桥接免疫测定有负面影响时的 ADA 检测。
