Functional Gold Nanoparticles-enhanced Electrochemical Detection of Glutathione Cross-linking Reaction and Sulfhydryl-specific Identification

Abstract

Abstract: An electrochemical detection method for the cross-linking reaction and sulfhydryl-specific identification of GSH using ferrocene-capped gold nanoparticles/streptavidin conjugates was developed in this paper. Disulfide C11-NHS organic molecules were covalently immobilized on a gold electrode via {Au-S} bonds(-S-S-is reduced to-SH easily in ethyl acetate solution) and formed compact self-assembled monolayers(SAMs). Then GSH is attached to C11-NHS through the amide formation and continues to cross-link Biotin-maleimide by forming a stable thioether bond between the sulfhydryl group and double bond of the maleimide group. Finally, the ferrocene-capped functional gold nanoparticles were introduced via strong interaction effect between biotin and streptavidin. The binding events were monitored by electrochemical signal of ferrocene covering on the gold nanoparticles by voltammetry. The signal differences permit to distinguish whether there is free sulfhydryl groups in the surface-bound biomolecules. The ferrocene signal is enlarged greatly because of quantities of ferrocene moieties covering on the nanogold surface. The well-defined redox peaks attribute to that of ferrocene immobilized through the cross-linking reaction between sulfhydryl groups in GSH and maleimide groups in Biotin-maleimide. The sensitivity of this electrochemical biosensor was detected and an excellent linearity of 5 μmol/L-0.1 mmol/L for GSH concentration with the oxidation current and a detection limit of 1 mmol/L were also achieved.

Key words: Glutathione, Functional gold nanoparticles, Ferrocene, Sulfhydryl-specific identification

CLC Number:

O657

TrendMD:

LI Jin-Hua, HU Jin-Bo, LI Qi-Long. Functional Gold Nanoparticles-enhanced Electrochemical Detection of Glutathione Cross-linking Reaction and Sulfhydryl-specific Identification[J]. Chem. J. Chinese Universities, 2005, 26(4): 647.

References

[1] CUI Yao-Zong(崔耀宗).Biochemistry(生物化学教程)[M],Hefei:Universityof Scienceand Technologyof China Press,1992 [2] Daniel H.M.,Peter H..J.Am.Chem.Soc.[J],2001,123:4062—4068 [3] Ellman G.L..Arch Biochem.Biophys.[J],1959,82:70—77 [4] Gleria K.D.,Hill H.A.O.,Wong L.L..FEBS Letters[J],1996,390:142—144 [5] Gleria K.D.,Halliwell C.M.,Jacob C.etal..FEBS Letters[J],1997,400:155—157 [6] Xu L.,Dong S..J.Electroanal.Chem.[J],1991,367:119—123 [7] Liu A.,Li T.,Wang E..Analyst[J],1995,11:597—601 [8] PEI Ren-Jun(裴仁军),CUI Xiao-Qiang(崔小强),YANG Xiu-Rong(杨秀荣)etal..Chem.J.Chinese Universities(高等学校化学学报)[J],2002,23(2):195—198 [9] Creager S.E.,Rowe G.K..J.Electroanal.Chem.[J],1994,370:203—211 [10] Hoogvliet J.C.,Dijksma M.,Kamp B.etal..Anal.Chem.[J],2000,70:2016—2021 [11] Wang J.,Li J.H.,Baca A.J.etal..Anal.Chem.[J],2003,75:3941—3945 [12] Baca A.J.,Zhou F.M.,Wang J.etal..Electroanalysis[J],2004,16(1—2):73—80 [13] WANG Xi-Cheng(王希成).Biochemistry(生物化学)[M],Beijing:Tsinghua University Press,2000: [14] XING Qi-Yi(邢其毅),XU Rui-Qin(徐瑞秋),ZHOU Zheng(周政)etal..Basic Organic Chemistry,Book 2nd.Ed.(基础有机化学,下册,第二版)[M],Beijing:Higher Education Press,1994:680—683 [15] Chidsey C.E.D.,Bertozzi C.R.,Putvinski T.M.etal..J.Am.Chem.Soc.[J],1990,112:4301—4309 [16] Bard A.J.,Faulkner L.R..Electrochemical Methods:Fundamentalsand Applications,2nd Ed.[M],New York:John Wileyand Sons,2001 [17] Senft A.P.,Dalon T.P.,Shertzer H.G..Analytical Biochemistry[J],2000,280(1):80—86 [18] Toyo′oka T.,Tanabe J.,Kashihara Y..Analytica Chimica Acta[J],2001,433(1):1—12 [19] Vander Jagt D.L.,Hunsaker L.A.,Vander Jagt T.J.etal..Biochemical Pharmacology[J],1997,53:1133—1140