Protein identification and quantification by two-dimensional infrared spectroscopy: implications for an all-optical proteomic platform.
Electron-vibration-vibration two-dimensional coherent spectroscopy, a variant of 2DIR, is shown to be a useful tool to differentiate a set of 10 proteins based on their amino acid content. Two-dimensional vibrational signatures of amino acid side chains are identified and the corresponding signal strengths used to quantify their levels by using a methyl vibrational feature as an internal reference. With the current apparatus, effective differentiation can be achieved in four to five minutes per protein, and our results suggest that this can be reduced to <1 min per protein by using the same technology. Finally, we show that absolute quantification of protein levels is relatively straightforward to achieve and discuss the potential of an all-optical high-throughput proteomic platform based on two-dimensional infrared spectroscopic measurements.
Fournier
Frédéric
F
Department of Chemistry, Imperial College London, Exhibition Road, London SW7 2AZ, United Kingdom.
Gardner
Elizabeth M
EM
Kedra
Darek A
DA
Donaldson
Paul M
PM
Guo
Rui
R
Butcher
Sarah A
SA
Gould
Ian R
IR
Willison
Keith R
KR
Klug
David R
DR
D000596:Amino Acids / Q000032:analysis / Q000737:chemistry; D055095:Optics and Photonics; D010449:Peptide Mapping / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity; D013055:Spectrophotometry, Infrared / Q000379:methods*
D000596:Amino Acids; D011506:Proteins