免疫去除血清高丰度蛋白对组织蛋白质组鉴定的影响

Impact of immunodepletion of serum high abundant protein on tissue proteome identification

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。

Objective To analyze the impact of immunodepletion of serum high abundant protein on identification of tis- sue proteome. Methods Human thyroid and pituitary samples were disposed using 12 high abundant protein depletion antibody column. The low abundant proteins and raw samples were analyzed by I_E/MS. Identified peptides and proteins were qualitatively and quantitatively analyzed. Results The serum high abundant proteins in thyroid and pituitary were depleted by 92. 11% ＋ 12. 87%, 78% ~25% respectively. Compared with raw thyroid, the non-redundant peptide and protein identification of low abundant thyroid were respectively increased by 70%, 16. 5%. The non-redundant peptide identification of low abundant pituitary was 21.9% higher than that of raw samples. But the protein identification was profile unchanged. Conclusions The immunedepletion of serum high abundant proteins can significantly improve pep- tide and protein identification and is more suitable for the tissues with intensive blood vessel.