辣根过氧化物酶信号放大法用于抗体覆盖的凝集素微阵列：提高靶向糖组分析灵敏度的策略。
Tyramide signal amplification for antibody-overlay lectin microarray: a strategy to improve the sensitivity of targeted glycan profiling.
机构信息
Department of Pathology, Johns Hopkins University, Baltimore, Maryland 21231, USA.
出版信息
J Proteome Res. 2011 Mar 4;10(3):1425-31. doi: 10.1021/pr1010873. Epub 2011 Jan 4.
Antibody-overlay lectin microarray (ALM) has been used for targeted glycan profiling to identify disease-related protein glycoforms. In this context, high sensitivity is desired because it allows for the identification of disease-related glycoforms that are often present at low concentrations. We describe a new tyramide signal amplification (TSA) for the antibody-overlay lectin microarray procedure for sensitive profiling of glycosylation patterns. We demonstrate that TSA increased the sensitivity of the microarray over 100 times for glycan profiling using the model protein prostate specific antigen (PSA). The glycan profile of PSA enriched from LNCAP cells, obtained at a subnanogram level with the aid of TSA, was consistent with the previous reports. We also established the glycan profile of prostate specific membrane antigen (PSMA) using the TSA and ALM. Thus, the TSA for antibody-overlay lectin microarray is a sensitive, rapid, comprehensive, and high-throughput method for targeted glycan profiling and can potentially be used for the identification of disease-related protein glycoforms.
抗体覆盖层凝集素微阵列（ALM）已被用于靶向糖组分析，以鉴定与疾病相关的蛋白糖型。在这种情况下，高灵敏度是理想的，因为它允许鉴定通常以低浓度存在的与疾病相关的糖型。我们描述了一种新的辣根过氧化物酶信号放大（TSA）用于抗体覆盖层凝集素微阵列程序，用于敏感的糖组分析。我们证明，TSA 使微阵列的灵敏度提高了 100 多倍，用于使用模型蛋白前列腺特异性抗原（PSA）的聚糖分析。使用 TSA 从 LNCAP 细胞中富集 PSA 的聚糖图谱与之前的报道一致。我们还使用 TSA 和 ALM 建立了前列腺特异性膜抗原（PSMA）的聚糖图谱。因此，抗体覆盖层凝集素微阵列的 TSA 是一种敏感、快速、全面和高通量的靶向聚糖分析方法，可用于鉴定与疾病相关的蛋白糖型。
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