title: Protein identification and quantification by two-dimensional infrared spectroscopy: implications for an all-optical proteomic platform.
abstract: 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.
authors.affiliation: Department of Chemistry, Imperial College London, Exhibition Road, London SW7 2AZ, United Kingdom.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Fournier
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Gardner
authors.forename: Darek A
authors.initials: DA
authors.lastname: Kedra
authors.forename: Paul M
authors.initials: PM
authors.lastname: Donaldson
authors.forename: Rui
authors.initials: R
authors.lastname: Guo
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Butcher
authors.forename: Ian R
authors.initials: IR
authors.lastname: Gould
authors.forename: Keith R
authors.initials: KR
authors.lastname: Willison
authors.forename: David R
authors.initials: DR
authors.lastname: Klug
mesh_terms: 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*
chemical_list: D000596:Amino Acids; D011506:Proteins