评估和优化 IgY 螺旋柱技术在人血清中丰富蛋白质去除中的应用。

Evaluation and optimization of IgY spin column technology in the depletion of abundant proteins from human serum.

机构信息

Division of Biology, Anatomy Department, Royal College of Surgeons in Ireland, Dublin, Ireland.

出版信息

Proteomics. 2011 Aug;11(16):3415-9. doi: 10.1002/pmic.201000587. Epub 2011 Jul 13.

DOI:10.1002/pmic.201000587

Abstract

Serum depletion strategies are commonly implemented in order to remove abundant proteins, increasing the number of proteins detected in a biomarker study. The IgY spin columns used in this study bind 12 and 14 primate proteins, respectively. 1-D SDS-PAGE and 2-DE revealed a suboptimal performance of the IgY spin columns. However, modification of the manufacturer's protocol, subjecting samples to two rounds of depletion, improved the number of proteins resolved by 2-DE. With alteration of the manufacturer protocol, the Seppro(®) IgY14 spin column can produce depleted serum with an increased number of spots resolved by 2-DE compared to untreated serum.

摘要

血清耗竭策略常用于去除丰富的蛋白质，从而增加生物标志物研究中检测到的蛋白质数量。本研究中使用的 IgY 螺旋柱分别结合了 12 种和 14 种灵长类蛋白质。1-D SDS-PAGE 和 2-DE 显示 IgY 螺旋柱的性能不佳。然而，修改制造商的方案，使样品经过两轮耗竭处理，可提高 2-DE 解析的蛋白质数量。通过修改制造商的方案，与未处理的血清相比，Seppro(®)IgY14 螺旋柱可以产生耗尽的血清，通过 2-DE 解析的斑点数量增加。