Selective metabolite and peptide capture/mass detection using fluorous affinity tags.
A new and general methodology is described for the targeted enrichment and subsequent direct mass spectrometric characterization of sample subsets bearing various chemical functionalities from highly complex mixtures of biological origin. Specifically, sample components containing a chemical moiety of interest are first selectively labeled with perfluoroalkyl groups, and the entire sample is then applied to a perfluoroalkyl-silylated porous silicon (pSi) surface. Due to the unique hydrophobic and lipophobic nature of the perfluorinated tags, unlabeled sample components are readily removed using simple surface washes, and the enriched sample fraction can then directly be analyzed by desorption/ionization on silicon mass spectrometry (DIOS-MS). Importantly, this fluorous-based enrichment methodology provides a single platform that is equally applicable to both peptide as well as small molecule focused applications. The utility of this technique is demonstrated by the enrichment and mass spectrometric analysis of both various peptide subsets from protein digests as well as amino acids from serum.
Go
Eden P
EP
Department of Molecular Biology and The Center for Mass Spectrometry, The Scripps Research Institute, La Jolla, California 92037, USA.
Uritboonthai
Wilasinee
W
Apon
Junefredo V
JV
Trauger
Sunia A
SA
Nordstrom
Anders
A
O'Maille
Grace
G
Brittain
Scott M
SM
Peters
Eric C
EC
Siuzdak
Gary
G
D000345:Affinity Labels / Q000737:chemistry*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000097:blood*; D006801:Humans; D006845:Hydrocarbons, Fluorinated / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D012825:Silicon / Q000737:chemistry; D013499:Surface Properties
D000345:Affinity Labels; D000596:Amino Acids; D006845:Hydrocarbons, Fluorinated; D010455:Peptides; D012825:Silicon