Improved analyte detectability of proteins and peptide lysates by means of multiple large-volume injection in LC-MS.
A 'multiple (trapping) large-volume injection' approach was developed for the analysis of peptides and proteins. In this way, a maximally 10-fold gain in sensitivity could be achieved. The system involves the use of an automated 10-port switching valve in combination with a 1 mm i.d. trapping/guard column and a 1 mm i.d. x 150 mm analytical column. The optimized multiple injection/loading procedure allows quantitative measurements of peptides and protein lysates. Linear calibration curves (R(2) > or = 0.988) over a minimum of two orders of magnitude were generated for a range of peptide and protein standards with sensitivities equal to or even exceeding, those generally achieved only through increasing miniaturization (quantification limit > or = 0.5 pmol/mL).
Storme
Michael L
ML
Laboratory of Medical Biochemistry and Clinical Analysis, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
t'Kindt
Ruben S
RS
Van Bocxlaer
Jan F
JF
D000818:Animals; D002138:Calibration; D002417:Cattle; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D055316:Cystatin C / Q000032:analysis*; D015891:Cystatins / Q000032:analysis*; D006454:Hemoglobins / Q000032:analysis*; D016014:Linear Models; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis*; D012680:Sensitivity and Specificity
D055316:Cystatin C; D015891:Cystatins; D006454:Hemoglobins; D010455:Peptides; C100158:cystatin, egg-white