• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Probing the binding kinetics of proinflammatory cytokine-antibody interactions using dual color fluorescence cross correlation spectroscopy
【24h】

Probing the binding kinetics of proinflammatory cytokine-antibody interactions using dual color fluorescence cross correlation spectroscopy

机译：使用双色荧光互相关光谱探测促炎细胞因子-抗体相互作用的结合动力学

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Dual color fluorescence cross correlation spectroscopy (FCCS) was used to investigate quantitatively the binding kinetics of tumor necrosis factor (TNFα) with TNFα antibody (anti-TNFα) following fluorescent labeling. Through the analysis of the auto correlation curves of fluorescence correlation spectroscopy (FCS), diffusion coefficients of 100.06 ± 4.9 μm~2 s~(-1) and 48.96 ± 2.52 μm~2 s~(-1) for Alexa488-TNFα and Atto647N-anti-TNFα were obtained. In addition, the calculated hydrodynamic diameters of the Alexa488-TNFα and Atto647N-anti-TNFα were approximately 4.89 ± 0.24 nm and 9.99 ± 0.52 nm, respectively, which agrees with the values of 5.20 ± 1.23 nm and 9.28 ± 0.86 nm for the native TNFα and the anti-TNFα as determined from dynamic light scattering measurements. For the binding kinetics, association (k_(on)) and dissociation (k_(off)) rate constants were (1.13 ± 0.08) × 10~4 M~(-1) s~(-1) and (1.53 ± 0.19) × 10~(-3) s~(-1)s while the corresponding dissociation constant (K_d) at 25 °C was (1.36 ± 0.10) × 10~(-7) M. We believe this is the first report on the binding kinetics for TNFα-antibody recognition in the homogeneous phase. Using this technology, we have shown that controlled experiments can be performed to gain insight into molecular mechanisms involved in the immune response.
机译：荧光标记后，使用双色荧光互相关光谱法（FCCS）定量研究肿瘤坏死因子（TNFα）与TNFα抗体（anti-TNFα）的结合动力学。通过分析荧光相关光谱（FCS）的自相关曲线，Alexa488-TNFα和Atto647N的扩散系数分别为100.06±4.9μm〜2 s〜（-1）和48.96±2.52μm〜2 s〜（-1）获得了抗TNFα。此外，Alexa488-TNFα和Atto647N-anti-TNFα的计算流体力学直径分别为约4.89±0.24 nm和9.99±0.52 nm，与天然值的5.20±1.23 nm和9.28±0.86 nm一致。由动态光散射测量确定的TNFα和抗TNFα。对于结合动力学，缔合（k_（on））和离解（k_（off））速率常数分别为（1.13±0.08）×10〜4 M〜（-1）s〜（-1）和（1.53±0.19） ×10〜（-3）s〜（-1）s，而在25°C时相应的解离常数（K_d）为（1.36±0.10）×10〜（-7）M。我们相信这是关于均相中TNFα-抗体识别的结合动力学。使用这项技术，我们已经表明可以进行受控实验以深入了解免疫应答中涉及的分子机制。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号