Sp3, single-pot, solid-phase-enhanced sample-preparation樣本處理方法經(jīng)典文獻(xiàn)參考
SP3(single-pot, solid-phase-enhanced sample-preparation)是一種基于順磁顆粒的蛋白質(zhì)組預(yù)處理工作流程,通過有機(jī)溶劑乙醇或乙腈驅(qū)動(dòng)驅(qū)動(dòng)蛋白在羧基或其它親水性磁珠表面被捕獲(如圖所示)。標(biāo)準(zhǔn) SP3流程中,蛋白質(zhì)通過親水相互作用機(jī)制與磁珠結(jié)合,結(jié)合了蛋白的磁珠被束縛在溶劑化層,可沖洗去除不需要的污染物,然后在一定條件下洗脫純化的蛋白質(zhì)。這種親水捕獲機(jī)制類似于親水相互作用色譜(HILIC)中所涉及的機(jī)制,已被廣泛證明是分離多種生物分子的有效工具。廈門普睿邁格生物科技有限公司開發(fā)的SP3-Carboxyl磁珠http://www.5000js.com/Product/1736805216.html可以完美替代SP3方案中最常使用的是Cytiva的Sera Mag SpeedBeads(GE Healthcare),提供了測定性能與成本之間最佳平衡。
以下是Sp3磁珠在蛋白組學(xué)樣本前處理方面的經(jīng)典文獻(xiàn),供參考:
- Hughes C S, Foehr S, Garfield D A, et al. Ultrasensitive proteome analysis using paramagnetic bead technology[J]. Molecular systems biology, 2014, 10(10): 757.
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Sielaff M, Kuharev J, Bohn T, et al. Evaluation of FASP, SP3, and iST protocols for proteomic sample preparation in the low microgram range[J]. Journal of proteome research, 2017, 16(11): 4060-4072.
- Moggridge S, Sorensen P H, Morin G B, et al. Extending the compatibility of the SP3 paramagnetic bead processing approach for proteomics[J]. Journal of proteome research, 2018, 17(4): 1730-1740.
- Hughes C S, Moggridge S, Müller T, et al. Single-pot, solid-phase-enhanced sample preparation for proteomics experiments[J]. Nature protocols, 2019, 14(1): 68-85.
- Hughes C S, Sorensen P H, Morin G B. A standardized and reproducible proteomics protocol for bottom-up quantitative analysis of protein samples using SP3 and mass spectrometry[J]. Proteomics for Biomarker Discovery: Methods and Protocols, 2019: 65-87.
- Müller T, Kalxdorf M, Longuespée R, et al. Automated sample preparation with SP 3 for low‐input clinical proteomics[J]. Molecular systems biology, 2020, 16(1): e9111.
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Mikulá?ek K, Kone?ná H, Potě?il D, et al. SP3 protocol for proteomic plant sample preparation prior LC-MS/MS[J]. Frontiers in plant science, 2021, 12: 635550.
- Duong V A, Lee H. Bottom-up proteomics: advancements in sample preparation[J]. International Journal of Molecular Sciences, 2023, 24(6): 5350.
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