Anal Chem. 2010 Dec 1;82(23):9601-5. doi: 10.1021/ac102241k. Epub 2010 Nov 10.
In this study, we report a new immunoassay platform using yeast cell surface display. This method holds promise for very low limit of detection (LOD) and is suitable for 2-Plex antibody recognition. Instead of adopting a conventional enzyme linked immunosorbent assay (ELISA) protocol by detecting the enzymatic activities or other physicochemical properties of the labeled analytes, this approach determines the quantity of an antibody analyte by directly counting the amount of "modified" yeast cells bound with antibody on the cell surface. c-myc and hemagglutinin (HA) tags were employed as an epitope model to demonstrate our approach. This yeast surface display based cell counting immunoassay (abbreviated as YSD-CCI) for anti-c-myc has a detection limit of 0.2 ng/mL, which is about 80 times higher than that of a conventional yeast ELISA under a similar condition. Moreover, the YSD-CCI's capability for 2-Plex antibody detection was demonstrated by simultaneous detection of anti-c-myc and anti-HA using engineered yeast cells expressing intracellular enhanced green fluorescent protein (EGFP) and mCherry, respectively. This proof-of-concept study paves the way for a new ultrasensitive multiplexed immunoassay method for diagnostic applications.
在本研究中，我们报告了一种使用酵母细胞表面展示的新免疫分析平台。该方法具有非常低的检测限（LOD）潜力，适合 2-Plex 抗体识别。与通过检测标记分析物的酶活性或其他物理化学性质的传统酶联免疫吸附测定（ELISA）方案不同，该方法通过直接计数与细胞表面抗体结合的“修饰”酵母细胞的数量来确定抗体分析物的数量。c-myc 和血凝素（HA）标签被用作表位模型来证明我们的方法。这种基于酵母表面展示的细胞计数免疫分析（简称 YSD-CCI）检测抗 c-myc 的检测限为 0.2ng/mL，在类似条件下，比传统的酵母 ELISA 高约 80 倍。此外，通过同时检测表达细胞内增强型绿色荧光蛋白（EGFP）和 mCherry 的工程化酵母细胞中的抗 c-myc 和抗 HA，证明了 YSD-CCI 用于 2-Plex 抗体检测的能力。这项概念验证研究为用于诊断应用的新型超灵敏多重免疫分析方法铺平了道路。
