基础医学与临床 ›› 2015, Vol. 35 ›› Issue (4): 439-443.

• 研究论文 • 上一篇    下一篇

免疫去除血清高丰度蛋白对组织蛋白质组鉴定的影响

邹丽莉1,李文婷2,王丹琪1,王曌1,孙伟3   

  1. 1. 中国医学科学院 北京协和医学院 基础学院
    2. 中国医学科学院基础医学研究所
    3. 中国医学科学院 北京协和医学院 基础医学研究所
  • 收稿日期:2014-12-26 修回日期:2015-01-14 出版日期:2015-04-05 发布日期:2015-04-08
  • 通讯作者: 孙伟 E-mail:sunwei1018@sina.com
  • 基金资助:
    国家重点基础研究发展计划;国家重点基础研究发展计划;中国科技部重点基础研究计划

Impact of serum high abundant protein immunodepletion on tissue proteome identification

  • Received:2014-12-26 Revised:2015-01-14 Online:2015-04-05 Published:2015-04-08

摘要: 目的 分析去除组织中血清高丰度蛋白对组织蛋白质组鉴定的影响。方法 用去除12种血清高丰度蛋白抗体柱处理人甲状腺和垂体组织样品,所得到的低丰度蛋白和原样分别进行液质联用分析,分别进行多肽和蛋白质水平的定性定量分析。结果 甲状腺和垂体组织中血清高丰度蛋白分别被除去了92.11%±12.87%和78%±25%。所鉴定到的多肽和蛋白与原样品相比,甲状腺组织的非冗余性多肽提高了70%、蛋白数目提高了16.5%,垂体的非冗余性多肽提高了21.9%,鉴定蛋白数目无明显变化。结论 去除组织中的血清高丰度蛋白法可以显著的提高肽段和蛋白的鉴定效率,更适用于血管密集的样品。

关键词: 甲状腺蛋白质组,垂体蛋白质组,去除血清高丰度蛋白、液质联用分析

Abstract: Objective To analyze the impact of serum high abundant proteins immunodepletion on identification of tissue proteome. Methods Human thyroid and pituitary samples were disposed using 12 high abundant proteins depletion antibody column. The low abundant proteins and raw samples were analyzed by LC/MS. Identified peptides and proteins were qualitatively and quantitatively analyzed. Results The serum high abundant proteins in thyroid and pituitary were respectively depleted by 92.11%±12.87%, 78%±25%. Compared with raw thyroid, the non-redundant peptide and protein identification of low abundant thyroid was respectively increased by 70%, 16.5%. The non-redundant peptide identification of low abundant pituitary was 21.9% higher than raw samples. But the protein identification was unchanged. Conclusions The immunodepletion of serum high abundant proteins can significantly increase peptide and protein identification and is more suitable for the tissues with intensive blood vessel.

Key words: thyroid proteome, pituitary proteome, serum high abundant proteins immunodepletion, LC/MS analysis

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