Rapid extraction and structural characterization of biomolecules in agarose gels by laser desorption Fourier transform mass spectrometry.
A method originally developed for the extraction of biomolecules from agarose gel slices has been utilized as a rapid means of isolating biological compounds from gels for subsequent structural characterization by matrix-assisted last desorption-ionization Fourier transform mass spectrometry (MALDI/FTMS). This "freeze-squeeze" extraction method involves pressure extrusion of fluid from frozen gel slices and provides near 50% recovery of analyte in less than 5 min. Experiments were directed at examining the recovery efficiency of the extraction method using 14C-labeled adenosine monophosphate and investigating the effect of high buffer concentrations on the laser desorption mass spectra. When coupled with this extraction technique, MALDI/FTMS can be used to detect and identify biomolecules at the low picomole level in agarose gel slices. The accurate mass measurements and MS/MS capabilities of the FTMS were exploited to provide detailed structural information at the isomeric level for oligonucleotides electrophoresed into agarose gels.
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta 30332-0400.
J C
JC
Dunphy
K L
KL
Busch
M V
MV
Buchanan
D000249:Adenosine Monophosphate / Q000737:chemistry / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005583:Fourier Analysis; D013058:Mass Spectrometry; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000302:isolation & purification*
D009838:Oligodeoxyribonucleotides; D000249:Adenosine Monophosphate