000081094 001__ 81094
000081094 005__ 20181203020303.0
000081094 0247_ $$2doi$$a10.1002/cphc.200390043
000081094 02470 $$2DAR$$a3772
000081094 02470 $$2ISI$$a000181790700004
000081094 037__ $$aARTICLE
000081094 245__ $$aReversible immobilization of peptides: Surface modification and in situ detection by attenuated total reflection FTIR spectroscopy
000081094 269__ $$a2003
000081094 260__ $$c2003
000081094 336__ $$aJournal Articles
000081094 520__ $$aA generic method is described for the reversible immobilization of polyhistidine-bearing polypeptides and proteins on attenuated total reflecting (ATR) sensor surfaces for the detection of biomol. interactions by FTIR spectroscopy. Nitrilotriacetic acid (NTA) groups are covalently attached to self-assembled monolayers of either thioalkanes on gold films or mercaptosilanes on silicon dioxide films deposited on germanium internal reflection elements. Complex formation between Ni2+ ions and NTA groups activates the ATR sensor surface for the selective binding of polyhistidine sequences. This approach not only allows a stable and reversible immobilization of histidine-tagged peptides (His-peptides) but also simultaneously allows the direct in situ quantification of surface-adsorbed mols. from their specific FTIR spectral bands. The surface concns. of both NTA and His-peptide on silanized surfaces were detd. to be 1.1 and 0.4 mols. nm-2, resp., which means that the surface is densely covered. A comparison of exptl. FTIR spectra with simulated spectra reveals a surface-enhancement effect of one order of magnitude for the gold surfaces. With the presented sensor surfaces, new ways are opened up to investigate, in situ and with high sensitivity and reproducibility, protein-ligand, protein-protein, protein-DNA interactions, and DNA hybridization by ATR-FTIR spectroscopy. [on SciFinder (R)]
000081094 700__ $$0241543$$aRigler, Per$$g113302
000081094 700__ $$0241544$$aUlrich, Wolf-Peter$$g111753
000081094 700__ $$0241019$$aHoffmann, Patrik$$g113150
000081094 700__ $$0241728$$aMayer, Michael$$g113417
000081094 700__ $$0240645$$aVogel, Horst$$g106666
000081094 773__ $$j4$$k3$$q268-275$$tChemPhysChem
000081094 909C0 $$0252153$$pLCPPM$$xU10170
000081094 909CO $$ooai:infoscience.tind.io:81094$$pSB$$particle
000081094 937__ $$aLCPPM-ARTICLE-2003-002
000081094 970__ $$a136/LCPPM
000081094 973__ $$aEPFL$$rREVIEWED$$sPUBLISHED
000081094 980__ $$aARTICLE