Pfizer, Inc, 558 Eastern Point Road Groton, CT 06340, United States.
J Immunol Methods. 2011 Aug 31;371(1-2):106-13. doi: 10.1016/j.jim.2011.06.020. Epub 2011 Jul 3.
Several detection platforms are available for ligand binding assays (LBA), each claiming superiority in sensitivity and dynamic range. However, little information exists in the literature directly comparing the various LBA platforms for quantitation. We have tested four common platforms to evaluate and compare the interchangeability of detection platforms by comparing sensitivity and dynamic range to a colorimetric LBA. The detection platforms compared are: colorimetric, chemiluminescence, time-resolved fluorescence (TRF) and electrochemiluminescence (ECL). Five different LBA protocols were tested with each of the detection endpoints. The assay protocols include the following ligand binding assay formats: direct binding, sandwich ELISA, competitive and cell based ELISA. We found that no detection platform consistently performed better than all the others and it was not possible to predict which platform would perform best for a given assay protocol. We also found surprising differences in assays (plate coating efficiency, low signal) which add to difficulty in choosing the best platform ad hoc. We propose here that in developing new assay protocols for detection of biotherapeutic agents, multiple detection platforms should be tested in order to forward the best assays possible and for the right reasons.
有几种检测平台可用于配体结合分析（LBA），每种平台都声称在灵敏度和动态范围方面具有优势。然而，文献中直接比较各种用于定量的 LBA 平台的信息很少。我们已经测试了四种常见的平台，通过将灵敏度和动态范围与比色 LBA 进行比较来评估和比较检测平台的可互换性。比较的检测平台有：比色法、化学发光法、时间分辨荧光（TRF）和电化学发光（ECL）。用每个检测终点测试了五种不同的 LBA 方案。测定方案包括以下配体结合测定格式：直接结合、三明治 ELISA、竞争和基于细胞的 ELISA。我们发现，没有一种检测平台始终优于所有其他平台，也不可能预测哪种平台对特定的测定方案表现最佳。我们还发现，在测定中存在令人惊讶的差异（板涂层效率、低信号），这增加了选择最佳平台的难度。我们在此建议，在为检测生物治疗剂开发新的测定方案时，应该测试多种检测平台，以便尽可能地推进最佳测定方案，并出于正确的原因。
