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Study of MMLV RT-Binding with DNA using surface plasmon resonance biosensor

Acta Biochimica et Biophysica Sinica 2005年第9期37卷 634-642页

作者： Lei WU~1 Ming-Hui HUANG~2 Jian-Long ZHAO~(1*) Meng-Su YANG~~(1,2*) 1 Shanghai Institute of Microsystem and Information Technology,Chinese Academy of Sciences,Shanghai 200050,China 2 Department of Bioloy and Chemistry,City University of Hong Kong,Hong Kong,China Shanghai Institute of Microsystem and Information Technology Chinese Academy of SciencesShanghai 200050 China Department of Biology and Chemistry City University of Hong KongHong Kong China

surface plasmon resonance biosensor technique was used to study the binding of Moloneymurine leukemia virus reverse transcriptase without RNase H domain （MMLV RT-） with DNA in the ab-sence and in the presence of *** DNA substrates,including single-stranded DNA （ssDNA）,DNA template-primer （T-P） duplex and gapped DNA,were immobilized on the biosensor chip surface usingstreptavidin-biotin,and MMLV RT--DNA binding kinetics were analyzed by different *** RT-could bind with ssDNA and the binding was involved in conformation *** RT- binding DNA T-Pduplex and gapped DNA could be analyzed using the simple 1:1 Langmuir *** lack of RNase Hdomain reduced the affinity between MMLV RT- and T-P *** effects of RT inhibitors,includingefavirenz,nevirapine and quercetin,on the interaction between MMLV RT- and gapped DNA were analyzedaccording to recovered kinetics *** slightly interfered with the binding between RT andDNA and the affinity constant in the presence of the inhibitor (KA=1.21×106 M-1) was lower than in theabsence of the inhibitor (KA=4.61×106 M-1).Nevirapine induced relatively tight binding between RT andDNA and the affinity constant in the presence of the inhibitor (KA=1.47×107 M-1) was approximately threefolds higher than without nevirapine,mainly due to rapid association and slow ***,aflavonoid originating from plant which has previously shown strong inhibition of the activity of *** to have minimal effect on the RT-DNA binding.

关键词： surface plasmon resonance biosensor reverse transcriptase kinetics inhibitor

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Polystyrene Based SPR biosensor Chip for Use in Immunoassay

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Tsinghua Science and Technology 1999年第3期4卷 1495-1498页

作者：杨军隋森芳 Department of Biological Sciences and Biotecbnology Tsinghua University Beijing 100084

biosensors are widely used in immunoassay. The biosensor chip carries a receptor which is used in immunoassay and the chip properties have an important influence on the detecting sensitivity of the biosensor. This paper describes a polystyrene\|based biosensor chip developed and used as part of a surface plasmon resonance (SPR) biosensor. The SPR biosensor has a much higher detecting sensitivity than enzyme\|linked immunoserbent assay (ELISA).

关键词： surface plasmon resonance biosensor polystyrene surface plasmon resonance microscopy enzyme\|linked immunoserbent assay

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