用于表面等离子体共振成像测量的在金基底上制备带有聚（L-谷氨酸）单分子层的DNA微阵列。
Fabrication of DNA microarrays with poly(L-glutamic acid) monolayers on gold substrates for SPR imaging measurements.
作者信息
Chen Yulin, Nguyen Anh, Niu Lifang, Corn Robert M
机构信息
Department of Chemistry, University of California--Irvine, Irvine, California 92697, USA.
出版信息
Langmuir. 2009 May 5;25(9):5054-60. doi: 10.1021/la804021t.
Abstract
Robust single-stranded DNA (ssDNA) microarrays are created by attaching amine-modified oligonucleotides to a monolayer of poly(L-glutamic acid) (pGlu) that is electrostatically adsorbed onto a chemically modified gold thin film. This surface attachment chemistry methodology is first characterized with a combination of polarization-modulation Fourier transform infrared (PM-FTIR) spectroscopy and surface plasmon resonance (SPR) angle shift measurements. SPR imaging (SPRI) measurements of these ssDNA microarrays are then used to study two surface bioaffinity interactions: (i) the quantitative hybridization adsorption of complementary ssDNA onto mixed ssDNA microarray elements and (ii) the adsorption of single-stranded binding protein (SSB) onto fully and partially hybridized DNA microarray elements. The Langmuir adsorption coefficient (K(Ads)) of SSB binding to ssDNA was determined to be (5.5 +/- 0.4) x 10(9) M(-1).
摘要
通过将胺修饰的寡核苷酸连接到静电吸附在化学修饰金薄膜上的聚（L-谷氨酸）（pGlu）单层上，制备出了坚固的单链DNA（ssDNA）微阵列。这种表面连接化学方法首先通过偏振调制傅里叶变换红外（PM-FTIR）光谱和表面等离子体共振（SPR）角位移测量相结合进行表征。然后，利用这些ssDNA微阵列的SPR成像（SPRI）测量来研究两种表面生物亲和相互作用：（i）互补ssDNA在混合ssDNA微阵列元件上的定量杂交吸附，以及（ii）单链结合蛋白（SSB）在完全和部分杂交的DNA微阵列元件上的吸附。确定SSB与ssDNA结合的朗缪尔吸附系数（K(Ads)）为(5.5 +/- 0.4) x 10(9) M(-1)。
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