Jung Kwanyoung, Cho Wonryeon, Regnier Fred E
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, USA.
J Proteome Res. 2009 Feb;8(2):643-50. doi: 10.1021/pr8007495.
Lectin affinity chromatography using concanavalin A (Con A), Helix pomatia agglutinin (HPA), Lycopersicon esculentum lectin (LEL), Aleuria aurantia lectin (AAL) and Lens culinaris agglutinin (LCA) was used to investigate the utility of narrow selectivity lectins in the characterization of plasma glycoproteome diversity and to recognize cancer associated aberrations in glycosylation. Following affinity chromatographic selection, proteins were tryptically digested, the peptide fragments separated by reversed phase chromatography (RPC), and fractions from RPC identified by tandem mass spectrometry. The diversity of glycosylation found with narrow selectivity lectins was generally 2/3 that of Con A and not related to protein abundance. Small groups of proteins were found with each of the affinity columns, HPA, LEL, AAL, and LCA, that changed 3-fold or more in concentration between normal and breast cancer patient plasma. Although the number of cancer patients examined was too small to validate cancer marker candidates, they are clearly worth examining in a larger, more diverse patient population.
使用伴刀豆球蛋白A（Con A）、蜗牛凝集素（HPA）、番茄凝集素（LEL）、橙黄银耳凝集素（AAL）和菜豆凝集素（LCA）进行凝集素亲和色谱分析，以研究窄选择性凝集素在表征血浆糖蛋白组多样性以及识别癌症相关糖基化异常方面的效用。经过亲和色谱筛选后，对蛋白质进行胰蛋白酶消化，通过反相色谱（RPC）分离肽片段，并通过串联质谱鉴定RPC的馏分。窄选择性凝集素发现的糖基化多样性通常是Con A的2/3，且与蛋白质丰度无关。在每个亲和柱（HPA、LEL、AAL和LCA）上都发现了一小部分蛋白质，它们在正常血浆和乳腺癌患者血浆之间的浓度变化了3倍或更多。尽管所检查的癌症患者数量太少，无法验证癌症标志物候选物，但它们显然值得在更大、更多样化的患者群体中进行检查。
