Development of a high-throughput IMS-IMS-MS approach for analyzing mixtures of biomolecules.
A high-throughput approach for biomolecule analysis is demonstrated for a mixture of peptides from tryptic digest of four proteins as well as a tryptic digests of human plasma. In this method a chip based electrospray autosampler coupled to a hybrid ion mobility (IMS) mass spectrometer (MS) is utilized to achieve rapid sample analysis. This high-throughput measurement is realized by exploiting the direct infusion capability of the chip based electrospray with its rapid sample manipulating capability as well as a high sensitive IMS-MS with a recently developed IMS-IMS separation technique that can be multiplexed to provide greater throughput. From replicate IMS-MS runs of known mixtures, the average uncertainty of peak intensities is determined to be +/-7% (relative standard deviation), and a detection limit in the low attomole range is established. The method is illustrated by analyzing 124 human plasma protein samples in duplicate, a measurement that required 16.5 h. Current limitations as well as implications of the high-throughput approach for complex biological sample analysis are discussed.
Kurulugama
Ruwan T
RT
Department of Chemistry, Indiana University, Bloomington, IN 47405, USA.
Valentine
Stephen J
SJ
Sowell
Rena A
RA
Clemmer
David E
DE
D016632:Apolipoprotein A-I / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry*; D004867:Equipment Design; D005656:Fungal Proteins / Q000737:chemistry; D006801:Humans; D007477:Ions; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013997:Time Factors; D014357:Trypsin / Q000737:chemistry*
D016632:Apolipoprotein A-I; D001798:Blood Proteins; D005656:Fungal Proteins; D007477:Ions; D010455:Peptides; D014357:Trypsin