Efficient enrichment and desalting of protein digests using magnetic mesocellular carbon foams in matrix-assisted laser desorption/ionization mass spectrometry.
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.
Kim
Young-Pil
YP
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea.
Cho
Kun
K
Lee
Dohoon
D
Piao
Yunxian
Y
Ahn
Yeong Hee
YH
Yoo
Jong Shin
JS
Hyun
Taeghwan
T
Kim
Hak-Sung
HS
D000803:Angiotensin I / Q000737:chemistry; D000818:Animals; D002384:Catalysis; D002417:Cattle; D008280:Magnetics; D053758:Nanoparticles / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D010449:Peptide Mapping; D016062:Porosity; D012235:Rhodamines / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013499:Surface Properties; D014357:Trypsin
C114557:5-carboxytetramethylrhodamine succinimidyl ester; D010446:Peptide Fragments; D012235:Rhodamines; D000803:Angiotensin I; D014357:Trypsin