Allosteric molecular sensing of anti-HIV antibodies by an immobilized engineered β-galactosidase

  • Ferraz R
  • Aris A
  • González G
 et al. 
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Abstract

The main objective is to demonstrate the feasibility of qualitative and semi-quantitative determination of anti-HIV antibodies, based on the allosteric response of an immobilized engineered Escherichia coli β-galactosidase. The suitability of this enzyme and the conditions for homogeneous assay had been previously demonstrated. In the present work, the operational conditions for engineered E. coli β-galactosidase immobilization on activated agarose supports have been determined. A solid biocatalyst has been prepared and characterized in order to determine the activation degree of the immobilized enzyme in the presence of anti-HIV antibodies. The solid-phase biocatalyst has been shown to be able to modulate its activity and produce an unequivocal signal in presence of anti-HIV containing sera from different patients. Finally, the conceptual design of a possible assay based on the above methodology is proposed. © 2007 Elsevier Inc. All rights reserved.

Author-supplied keywords

  • Allosteric enzymes
  • Anti-HIV antibodies sensing
  • Immobilized engineered β-galactosidase

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Authors

  • Gregorio AlvaroUniversitat Autonoma de Barcelona Escola d'Enginyeria

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  • R. M. Ferraz

  • A. Aris

  • G. González

  • J. López-Santín

  • A. Villaverde

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