Kim Young-Pil, Cho Kun, Lee Dohoon, Piao Yunxian, Ahn Yeong Hee, Yoo Jong Shin, Hyun Taeghwan, Kim Hak-Sung
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea.
Rapid Commun Mass Spectrom. 2007;21(21):3435-42. doi: 10.1002/rcm.3231.
We demonstrate that magnetic mesocellular carbon foams (Mag-MCF-C) can be effectively used for enrichment and desalting of protein digests or peptides in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The large mesocellular pores and surface area of Mag-MCF-C are likely to mainly contribute to high efficiency in enrichment and desalting of protein digests. The magnetic property of Mag-MCF-C enabled easy and simple enrichment and desalting process comprising adsorption, washing, and separation steps by using an external magnet. Following elution from Mag-MCF-C by using a matrix solution (CHCA in 70% ACN/0.1% TFA), the peptides were subjected to MALDI-MS analysis. As a result, MALDI mass spectra of peptides or tryptic protein digests were distinct even at a peptide concentration as low as 50 pM. The use of Mag-MCF-C resulted in significantly improved sequence coverage for protein identification when compared to other conventional methods. Mag-MCF-C will find applications in mass spectrometric analysis of low abundance peptides or protein digests with high sensitivity.
我们证明,磁性介孔碳泡沫(Mag-MCF-C)可有效地用于基质辅助激光解吸/电离质谱(MALDI-MS)中蛋白质消化物或肽段的富集和脱盐。Mag-MCF-C的大介孔和表面积可能是蛋白质消化物高效富集和脱盐的主要原因。Mag-MCF-C的磁性使得通过使用外部磁铁进行包括吸附、洗涤和分离步骤的富集和脱盐过程变得简单易行。用基质溶液(70%乙腈/0.1%三氟乙酸中的CHCA)从Mag-MCF-C上洗脱后,对肽段进行MALDI-MS分析。结果,即使在肽段浓度低至50 pM时,肽段或胰蛋白酶消化蛋白质的MALDI质谱也很清晰。与其他传统方法相比,使用Mag-MCF-C可显著提高蛋白质鉴定的序列覆盖率。Mag-MCF-C将在低丰度肽段或蛋白质消化物的高灵敏度质谱分析中得到应用。