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受体介导的黏附现象。自由基流动分离测定法的模型研究。

Receptor-mediated adhesion phenomena. Model studies with the Radical-Flow Detachment Assay.

作者信息

Cozens-Roberts C, Quinn J A, Lauffenberger D A

机构信息

Department of Chemical Engineering, University of Pennsylvania, Philadelphia 19104.

出版信息

Biophys J. 1990 Jul;58(1):107-25. doi: 10.1016/S0006-3495(90)82357-2.

Abstract

Receptor-mediated cell adhesion phenomena play a vital role in many physiological and biotechnology-related processes. To investigate the physical and chemical factors that influence the cell/surface interaction, we have used a radial flow device, a so-called Radial-Flow Detachment Assay (RFDA). The RFDA allows us to make direct observations of the detachment process under specified experimental conditions. In results reported here, we have studied the detachment of receptor-coated latex beads (prototype cells) from ligand-coated glass surfaces. The receptors and ligands used in this work are complementary antibodies. The beads enable us to examine several aspects of the adhesion process with particles having uniform properties that can be varied systematically. Advantages of the RFDA are many, especially direct observation of cell detachment over a range of shear stresses with quantitative measurement of the adhesive force. We focus our studies on the effects of ligand and receptor densities, along with the influence of pH and ionic strength of the medium. These data are analyzed with a mathematical model based on the theoretical framework of Bell, G. I. (1978. Science [Wash. DC]. 200:618-627) and Hammer, D. A. and D. A. Lauffenburger (1987. Biophys. J. 52:475-487). We demonstrate experimental validation of a theoretical expression for the critical shear stress for particle detachment, and show that it is consistent with reasonable estimates for the receptor-ligand bond affinity.

摘要

受体介导的细胞黏附现象在许多生理和生物技术相关过程中起着至关重要的作用。为了研究影响细胞/表面相互作用的物理和化学因素,我们使用了一种径向流动装置,即所谓的径向流动分离测定法(RFDA)。RFDA使我们能够在特定实验条件下直接观察分离过程。在本文报道的结果中,我们研究了受体包被的乳胶珠(原型细胞)从配体包被的玻璃表面的分离。本研究中使用的受体和配体是互补抗体。这些珠子使我们能够用具有可系统变化的均匀性质的颗粒来研究黏附过程的几个方面。RFDA的优点很多,特别是能够在一系列剪切应力下直接观察细胞分离,并对黏附力进行定量测量。我们将研究重点放在配体和受体密度的影响以及介质pH值和离子强度的影响上。这些数据使用基于Bell, G. I.(1978年,《科学》[华盛顿特区],200:618 - 627)以及Hammer, D. A.和D. A. Lauffenburger(1987年,《生物物理杂志》,52:475 - 487)理论框架的数学模型进行分析。我们展示了颗粒分离临界剪切应力理论表达式的实验验证,并表明它与受体 - 配体键亲和力的合理估计值一致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7080/1280944/4ba9a7e512d6/biophysj00125-0116-a.jpg

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