Desorption electrospray ionization-mass spectrometry of proteins.
Desorption electrospray ionization-mass spectrometry (DESI-MS) was evaluated for the detection of proteins ranging in molecular mass from 12 to 66 kDa. Proteins were uniformly deposited on a solid surface without pretreatment and analyzed with a DESI source coupled to a quadrupole ion trap mass spectrometer. DESI-MS parameters optimized for protein detection included solvent flow rate, temperature of heated capillary tube, incident and reflection angle, sheath gas pressure, and ESI voltage. Detection limits were obtained for all protein standards, and they were found to decrease with decreasing protein molecular mass: for cytochrome c (12.3 kDa) and lysozyme (14.3 kDa) a detection limit of 4 ng/mm2 was obtained; for apomyoglobin (16.9 kDa) 20 ng/mm2; for beta-lactoglobulin B (18.2 kDa) 50 ng/mm2; and for chymotrypsinogen A (25.6 kDa) 100 ng/mm2. The DESI-MS analysis of higher molecular mass proteins such as ovalbumin (44.4 kDa) and bovine serum albumin (66.4 kDa) yielded mass spectra of low signal-to-noise ratio, making their detection and molecular weight determination difficult. In this study, DESI-MS proved to be a rapid and robust method for accurate MW determination for proteins up to 17 kDa under ambient conditions. Finally, we demonstrated the DESI-MS detection of the bacteriophage MS2 capsid protein from crude samples with minimal sample preparation.
Shin
Yong-Seung
YS
Department of Chemistry, Arthropod-Borne Animal Diseases Research Laboratory, ARS, USDA, and School of Pharmacy, University of Wyoming, 1000 East University Avenue, Laramie, Wyoming 82071, USA.
Drolet
Barbara
B
Mayer
Richard
R
Dolence
Kurt
K
Basile
Franco
F
D000818:Animals; D001059:Apoproteins / Q000032:analysis; D036022:Capsid Proteins / Q000032:analysis; D002417:Cattle; D002645:Chickens; D002919:Chymotrypsinogen / Q000032:analysis; D045304:Cytochromes c / Q000032:analysis; D004926:Escherichia coli / Q000737:chemistry; D007782:Lactoglobulins / Q000032:analysis; D017909:Levivirus / Q000737:chemistry; D008970:Molecular Weight; D009113:Muramidase / Q000032:analysis; D009211:Myoglobin / Q000032:analysis; D010047:Ovalbumin / Q000032:analysis; D010316:Particle Size; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D001059:Apoproteins; D036022:Capsid Proteins; D007782:Lactoglobulins; D009211:Myoglobin; D011506:Proteins; C010857:apomyoglobin; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D045304:Cytochromes c; D002919:Chymotrypsinogen; D009113:Muramidase