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通过 shotgun 蛋白质组学对七种商业化人血清增富/ depleted 产品的比较评估。

Comparative evaluation of seven commercial products for human serum enrichment/depletion by shotgun proteomics.

机构信息

Porto Conte Ricerche, Science and Technology Park of Sardinia, Tramariglio, Alghero, Sassari, Italy.

Porto Conte Ricerche, Science and Technology Park of Sardinia, Tramariglio, Alghero, Sassari, Italy.

出版信息

Talanta. 2018 Aug 1;185:213-220. doi: 10.1016/j.talanta.2018.03.086. Epub 2018 Mar 28.

Abstract

Seven commercial products for human serum depletion/enrichment were tested and compared by shotgun proteomics. Methods were based on four different capturing agents: antibodies (Qproteome Albumin/IgG Depletion kit, ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit, Top 2 Abundant Protein Depletion Spin Columns, and Top 12 Abundant Protein Depletion Spin Columns), specific ligands (Albumin/IgG Removal), mixture of antibodies and ligands (Albumin and IgG Depletion SpinTrap), and combinatorial peptide ligand libraries (ProteoMiner beads), respectively. All procedures, to a greater or lesser extent, allowed an increase of identified proteins. ProteoMiner beads provided the highest number of proteins; Albumin and IgG Depletion SpinTrap and ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit resulted the most efficient in albumin removal; Top 2 and Top 12 Abundant Protein Depletion Spin Columns decreased the overall immunoglobulin levels more than other procedures, whereas specifically gamma immunoglobulins were mostly removed by Albumin and IgG Depletion SpinTrap, ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit, and Top 2 Abundant Protein Depletion Spin Columns. Albumin/IgG Removal, a resin bound to a mixture of protein A and Cibacron Blue, behaved less efficiently than the other products.

摘要

七种商业化的人血清耗竭/富集产品通过鸟枪法蛋白质组学进行了测试和比较。方法基于四种不同的捕获剂:抗体(Qproteome Albumin/IgG Depletion kit、ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit、Top 2 Abundant Protein Depletion Spin Columns 和 Top 12 Abundant Protein Depletion Spin Columns)、特异性配体(Albumin/IgG Removal)、抗体和配体混合物(Albumin and IgG Depletion SpinTrap)以及组合肽配体文库(ProteoMiner beads)。所有这些方法在不同程度上都允许鉴定到更多的蛋白质。ProteoMiner beads 提供了最多的蛋白质;Albumin and IgG Depletion SpinTrap 和 ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit 在去除白蛋白方面最有效;Top 2 和 Top 12 Abundant Protein Depletion Spin Columns 比其他方法更能降低总免疫球蛋白水平,而 Albumin and IgG Depletion SpinTrap、ProteoPrep Immunoaffinity Albumin and IgG Depletion Kit 和 Top 2 Abundant Protein Depletion Spin Columns 则主要去除 γ 免疫球蛋白。Albumin/IgG Removal,一种与蛋白 A 和 Cibacron Blue 的混合物结合的树脂,其性能不如其他产品。

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