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Probing single molecule interactions by AFM using bio-functionalized dendritips

机译:使用生物功能化的树突尖通过AFM探测单分子相互作用

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摘要

A new strategy for AFM (atomic force microscopy) tip functionalization based on the use of aldehyde-phosphorus dendrimers for the immobilization of biomolecules such as proteins (e.g. antibodies) is presented. Firstly functionalized with amino groups, the tips are reacted with dendrimers leading to dendrimer-activated tips (so-called dendritips). Free aldehyde functions on the dendrimer are therefore available to react with amino-functions present on every protein and many biomolecules. Using biofunctionalized-dendritip, single molecule force interactions between glutathione-S-transferase (GST) and its cognate antibody immobilized on dendritips (67 ± 11 pN for single interaction) were probed by AFM spectroscopy. The specificity of our measurements was demonstrated by performing blocking tests resulting in the loss of interactions.
机译:提出了一种新的AFM(原子力显微镜)尖端功能化策略,该策略基于使用醛-磷树状大分子固定生物分子(例如蛋白质)(例如抗体)。首先用氨基官能化,使尖端与树枝状聚合物反应,从而导致树枝状聚合物活化的尖端(所谓的树枝状尖端)。因此,树枝状聚合物上的游离醛官能团可与存在于每种蛋白质和许多生物分子上的氨基官能团反应。使用生物功能化的树突状细胞,谷胱甘肽-S-转移酶(GST)与其固定在树突状细胞上的同源抗体(单次相互作用为67±11 pN)之间的单分子力相互作用通过原子力显微镜(AFM)光谱进行了探测。我们的测量的特异性通过进行封闭试验导致相互作用的丧失来证明。

著录项

  • 来源
    《Sensors and Actuators》 |2012年第2012期|p.436-441|共6页
  • 作者单位

    Universite de Toulouse, LPCNO, INSA, CNRS, UPS, 135 Av. de Rangueil, F-31077 Toulouse, France,CNRS, LAAS. 7 avenue du Colonel Roche, F-31077 Toulouse, France,Universite de Toulouse, UPS, INSA, 1NP, ISAE, UT1, UTM, LAAS, F-31077 Toulouse, France,CNRS, ITAV-UMS3039, F-31106 Toulouse, France,University de Toulouse, ITAV-UMS3039, F-31106 Toulouse, France;

    CNRS, LAAS. 7 avenue du Colonel Roche, F-31077 Toulouse, France,Universite de Toulouse, UPS, INSA, 1NP, ISAE, UT1, UTM, LAAS, F-31077 Toulouse, France,CNRS, ITAV-UMS3039, F-31106 Toulouse, France,University de Toulouse, ITAV-UMS3039, F-31106 Toulouse, France;

    Laboratoire de Chimie de Coordination du CNRS (LCC), BP 44099,205, route de Narbonne, F-31077 Toulouse Cedex 4, France,Universite de Toulouse, UPS, INPT, LCC, F-31077 Toulouse, France;

    Universite de Toulouse, LPCNO, INSA, CNRS, UPS, 135 Av. de Rangueil, F-31077 Toulouse, France;

    Laboratoire de Chimie de Coordination du CNRS (LCC), BP 44099,205, route de Narbonne, F-31077 Toulouse Cedex 4, France,Universite de Toulouse, UPS, INPT, LCC, F-31077 Toulouse, France;

    Laboratoire de Chimie de Coordination du CNRS (LCC), BP 44099,205, route de Narbonne, F-31077 Toulouse Cedex 4, France,Universite de Toulouse, UPS, INPT, LCC, F-31077 Toulouse, France;

    CNRS, ITAV-UMS3039, F-31106 Toulouse, France,University de Toulouse, ITAV-UMS3039, F-31106 Toulouse, France,Universite de Toulouse, INSA, UPS, INP, LISBP, 135 Av. de Rangueil, F-31077 Toulouse, France,INRA, UMR792 lngenierie des Systemes Biologiques et des Procedes, F-31400 Toulouse, France,Universite de Toulouse, LPCNO, INSA, CNRS, UPS, 135 Av. de Rangueil, F-31077 Toulouse, France;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    AFM tip functionalization; phosphorus dendrimers; single molecule force spectroscopy (SMFS); molecular recognition; antigen/antibody interaction;

    机译:AFM尖端功能化;磷树枝状聚合物单分子力谱(SMFS);分子识别抗原/抗体相互作用;

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