Acid cleavable surface enhanced raman tagging for protein detection.
Dye conjugation is a common strategy improving the surface enhanced Raman detection sensitivity of biomolecules. Reported is a proof-of-concept study of a novel surface enhanced Raman spectroscopic tagging strategy termed as acid-cleavable SERS tag (ACST) method. Using Rhodamine B as the starting material, we prepared the first ACST prototype that consisted of, from the distal end, a SERS tag moiety (STM), an acid-cleavable linker, and a protein reactive moiety. Complete acid cleavage of the ACST tags was achieved at a very mild condition that is 1.5% trifluoroacetic acid (TFA) aqueous solution at room temperature. SERS detection of this ACST tagged protein was demonstrated using bovine serum albumin (BSA) as the model protein. While the SERS spectrum of intact ACST-BSA was entirely dominated by the fluorescent signal of STM, quality SERS spectra can be readily obtained with the acid cleaved ACST-BSA conjugates. Separation of the acid cleaved STM from protein further enhances the SERS sensitivity. Current SERS detection sensitivity, achieved with the acid cleaved ACST-BSA conjugate is ∼5 nM in terms of the BSA concentration and ∼1.5 nM in ACST content. The dynamic range of the cleaved ACST-BSA conjugate spans four orders of magnitudes from ∼10 nM to ∼100 μM in protein concentrations. Further improvement in the SERS sensitivity can be achieved with resonance Raman acquisition. This cleavable tagging strategy may also be used for elimination of protein interference in fluorescence based biomolecule detection.
Department of Chemistry, Mississippi State University, Mississippi State, MS 39762, USA. dz33@msstate.edu
Dongmao
D
Zhang
Karthikeshwar
K
Vangala
Shaoyong
S
Li
Michael
M
Yanney
Hao
H
Xia
Sige
S
Zou
Andrzej
A
Sygula
D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry*; D015394:Molecular Structure; D011506:Proteins / Q000032:analysis*; D012235:Rhodamines / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013379:Substrate Specificity; D014269:Trifluoroacetic Acid / Q000737:chemistry*
D005456:Fluorescent Dyes; D011506:Proteins; D012235:Rhodamines; D012710:Serum Albumin, Bovine; D014269:Trifluoroacetic Acid; C029773:rhodamine B