Determination of proteins in complex biological matrices has massive attention and is exploited in many different scientific disciplines. Routinely, proteins are determined using antibody based immuno-metric assays. Although these assays are easy to perform and widely used, interpretation of the results is challenging: cross reactivity, high dose hook-effect, presence of heterophile- or auto-antibodies give rise to false results, sometimes with dramatic consequences. In the quest for more robust assays a combination of antibody sample clean-up, tryptic digestion and mass spectrometric determination is gaining more attention. This review discusses the advantages of antibody based affinity capture and subsequent LC-MS/MS in protein analysis like less false results and possibilities like multiplexing and isoform differentiation. It also considers the interplay between the analytical, biological and biochemical factors, which still give rise to false results, even with mass spectrometry as the ultimate selective detection step. The intention of this review is to point out both strengths and weaknesses of antibody based affinity capture LC-MS/MS in quantitative determination of proteins in biological matrices.

中文翻译：

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基于抗体的亲和捕获LC-MS / MS用于定量测定生物基质中的蛋白质

复杂生物基质中蛋白质的测定受到了广泛关注，并在许多不同的科学学科中得到了广泛应用。通常，使用基于抗体的免疫测定法确定蛋白质。尽管这些测定法易于执行且用途广泛，但对结果的解释却具有挑战性：交叉反应性，高剂量的钩连效应，存在嗜异性或自体抗体会产生错误的结果，有时会产生严重的后果。为了寻求更强大的分析方法，抗体样品净化，胰蛋白酶消化和质谱测定的结合越来越受到关注。这篇综述讨论了基于抗体的亲和捕获和随后的LC-MS / MS在蛋白质分析中的优势，如较少的错误结果以及诸如多重分析和同工型分化的可能性。它还考虑了分析，生物和生化因素之间的相互作用，即使质谱作为最终的选择性检测步骤，这些相互作用仍然会导致错误的结果。这篇综述的目的是指出基于抗体的亲和捕获LC-MS / MS在定量测定生物基质中蛋白质时的优点和缺点。