Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, Northern Ireland, UK.
LGC, Teddington, UK.
Compr Rev Food Sci Food Saf. 2023 Sep;22(5):3984-4003. doi: 10.1111/1541-4337.13216. Epub 2023 Aug 2.
Food allergy remains a public health, business, and regulatory challenge. Risk analysis (RA) and risk management (RM) of food allergens are of great importance and analysis for food allergens is necessary for both. The current workhorse techniques for allergen analysis (enzyme linked immunosorbent assay [ELISA] and real-time polymerase chain reaction) exhibit recognized challenges including variable and antibody specific responses and detection of species DNA rather than allergen protein, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) enables protein identification, with potential for multiplex analysis and traceability to the System of International units (SI), aiding global measurement standardization. In this review, recent literature has been systematically reviewed to assess progress in LC-MS/MS and define the potential and benefits of matrix-assisted laser desorption/ionization-time-of-flight MS (MALDI-ToF-MS) technology for allergen analysis. MALDI-ToF-MS of initially intact protein is already applied to verify in silico-derived peptide sequences for LC-MS/MS analysis. We describe the origins of MALDI and its future perspectives, including affinity bead-assisted assays coupled to MALDI. Based on the proliferation of reliable and reproducible MALDI-based clinical applications, the technique should emulate the detection capability (sensitivity) of established allergen detection techniques, whilst reducing technical support and having equivalent multiplexing potential to competing techniques, for example, LC-MS/MS and ELISA. Although unlikely to offer inherent SI traceability, MALDI-based allergen analysis will complement existing MS approaches for allergens. Affinity bead-MALDI appears capable of higher throughput at lower cost per sample than almost any existing technique, enabling repeated sub-sampling as a way to reduce representative sampling issues.
食物过敏仍然是一个公共卫生、商业和监管方面的挑战。食物过敏原的风险分析（RA）和风险管理（RM）非常重要，两者都需要对食物过敏原进行分析。目前用于过敏原分析的主要技术（酶联免疫吸附测定[ELISA]和实时聚合酶链反应）分别存在着可变性和抗体特异性反应以及检测物种 DNA 而不是过敏原蛋白等公认的挑战。液相色谱-串联质谱（LC-MS/MS）能够进行蛋白质鉴定，具有多重分析和可追溯到国际单位制（SI）的潜力，有助于实现全球测量标准化。在这篇综述中，我们系统地回顾了最近的文献，以评估 LC-MS/MS 的进展，并定义基质辅助激光解吸/电离-飞行时间质谱（MALDI-ToF-MS）技术在过敏原分析中的潜力和优势。最初完整蛋白质的 MALDI-ToF-MS 已经用于验证 LC-MS/MS 分析的基于计算机的肽序列。我们描述了 MALDI 的起源及其未来展望，包括与 MALDI 偶联的亲和珠辅助测定。基于可靠和可重复的 MALDI 临床应用的激增，该技术应该模仿已建立的过敏原检测技术的检测能力（灵敏度），同时降低技术支持，并具有与竞争技术（例如 LC-MS/MS 和 ELISA）相当的多重化潜力。尽管不太可能提供固有的 SI 可追溯性，但基于 MALDI 的过敏原分析将补充现有的 MS 方法用于过敏原。亲和珠-MALDI 似乎能够以比几乎任何现有技术更低的成本实现更高的通量，通过重复亚采样来降低代表性采样问题。
