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Improvement of homogeneity of analytical biodevices by gene manipulation
Shi, JX; Zhang, XE; Xie, WH; Zhou, YF; Zhang, ZP; Deng, JY; Cass, AEG; Zhang, ZL; Pang, DW; Zhang, CG
Corresponding AuthorZhang, XE()
2004-02-01
Source PublicationANALYTICAL CHEMISTRY
ISSN0003-2700
Volume76Issue:3Pages:632-638
AbstractHomogeneity is proposed for evaluation of the quality of analytical biodevices, such as biosensors and biochips. As a demonstration, glucose oxidase (GOx) was modified at its C-terminal with a linker peptide with a cysteine residue at the end. The fusion structure (GOx-linker-cysteine) enables the enzyme to immobilize on gold surfaces with a Cys-S-Au bond or to immobilize on a silanized glass surface via disulfide chemistry. With this fusion structure, the enzyme can be anchored onto the substrate with well-controlled orientation, thus forming a homogeneous biological layer on biodevices. The linker peptide between GOx and the cysteine acts as a spacer to reduce the steric hindrance caused by the bulky body of the enzyme. Biochemistry experiments showed that this genetically modified glucose oxidase (shortened to GOxm) retained most of its catalytic characteristics, with Km and,at similar to those of the wild-type GOx. Electrochem, istry experiments showed that GOxm-modified electrode gave higher and more stable current responses than the electrode modified with GOx which has no free -SH on its surface. The coefficients of variation. (used for evaluation of the interchangeability of the enzyme device from the same batch preparation) were 9.5% for the GOxm gold electrode and 20.0% for the GOx gold electrode and the GOxm oxygen electrode. The relative errors (used for evaluation of the precision of the individual enzyme device) were 2.9% for the GOxm gold electrode, 12.0% for the GOx gold electrode, and 11.2% for the GOxm oxygen electrode. Atomic force microscopy images revealed that GOxm formed a self-assembled monolayer in a hexagonal-like lattice packing arrangement on the gold surface, while GOx formed multilayer assembling or aggregated particles. The homogeneity of the protein chips, the GOxm array that was prepared through -S-S- formation, and the GOx array that was prepared through nonspecific adsorption was evaluated. The coefficients of variation, calculated with the signal level of all dots, were 5.4% for the GOxm array and 81.8% for the GOx array. All experimental results pointed to the fact that the homogeneity of the analytical biodevices could be considerably improved by using the proposed method.
DOI10.1021/ac020796f
Indexed BySCI
Language英语
WOS Research AreaChemistry
WOS SubjectChemistry, Analytical
WOS IDWOS:000188823900024
PublisherAMER CHEMICAL SOC
Citation statistics
Cited Times:17[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.imr.ac.cn/handle/321006/126067
Collection中国科学院金属研究所
Corresponding AuthorZhang, XE
Affiliation1.Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China
2.Chinese Acad Sci, Shenyang Inst Ecol, Shenyang, Peoples R China
3.Wuhan Univ, Dept Chem, Wuhan 430072, Peoples R China
4.Univ London Imperial Coll Sci Technol & Med, Dept Biochem, London SW7 2AY, England
Recommended Citation
GB/T 7714
Shi, JX,Zhang, XE,Xie, WH,et al. Improvement of homogeneity of analytical biodevices by gene manipulation[J]. ANALYTICAL CHEMISTRY,2004,76(3):632-638.
APA Shi, JX.,Zhang, XE.,Xie, WH.,Zhou, YF.,Zhang, ZP.,...&Zhang, CG.(2004).Improvement of homogeneity of analytical biodevices by gene manipulation.ANALYTICAL CHEMISTRY,76(3),632-638.
MLA Shi, JX,et al."Improvement of homogeneity of analytical biodevices by gene manipulation".ANALYTICAL CHEMISTRY 76.3(2004):632-638.
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