摘要
研究了基于亲和型生物传感器的白细胞介素-1α(IL-1α)与I型可溶性白细胞介素-1受体(sIL-1RⅠ)相互作用的实时监测,并应用FASTfit软件对数据进行拟合分析。采用共价交联法在样品池表面固定sIL-1RⅠ,其密度为2.13 ng/mm2,用于检测不同浓度IL-1α与sIL-1RⅠ的结合所引起响应信号变化。对浓度为2400 nmol/L IL-1α的响应信号为122.66弧度秒,并且响应信号同IL-1α浓度呈线性关系,相关系数为0.972。FASTfit拟合数据结果表明IL-1α与sIL-1RⅠ相互作用符合一级动力学方程,其误差值不超过0.5弧度秒,其解离平衡常数(KD)和结合平衡常数(KA)分别为4.15×10-6mol/L和2.41×105mol/L。验证了亲和型生物传感器研究IL-1α和sIL-1RⅡ相互作用及动力学分析的可行性,有助于阐明IL-1α和sIL-1RⅠ的重要生物学功能,并为快速筛选其拮抗剂研究提供了理论和实验数据。
An affinity biosensor based on the resonant mirror was employed to monitor the interaction and kinetic analysis of Interleukin-1α （IL-1α） and type Ⅰ soluble Intefleukin-1 receptor （ sIL-1R Ⅰ ） , and the association data between IL-1α and sIL-1R Ⅰ was fitted by FASTfit software, sIL-1R Ⅰ was coupled to carboxymethyl dextran （CMD） cuvette surface by the covalent effect of amine to set up the experimental model, with a density of 2. 13 ng/mm^2, which was used to bind to IL-1α in solution. For 2400 nmol/L IL-1α, the response was about 122.66 arc seconds. For the different concentrations of IL-1α, the responses were linear to the concentrations and the linear correlation coefficient was 0.9972. The binding curve was fitted by FASTfit analysis, which fitted the monophasic association quite well, and the error did not exceed 0.5 arc seconds. For the interaction, KA was 2.41 × 10^5 （mol/L） ^-1, and KD was 4.15 × 10^-6 mol/L. The experiment verifies the feasibility of using an affinity biosensor to real time monitor the interaction and kinetic analysis of IL-1α and sIL-1R Ⅰ. All these not only can help to clarify the biological function of IL-1α and sIL-1R Ⅰ, but also can provide principle and experiment data for the identification and filtration of their effective antagonists.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2006年第6期769-772,共4页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金资助项目(No.90209054)
