TY - JOUR
T1 - Allosteric molecular sensing of anti-HIV antibodies by an immobilized engineered β-galactosidase
AU - Ferraz, R. M.
AU - Aris, A.
AU - González, G.
AU - López-Santín, J.
AU - Villaverde, A.
AU - Álvaro, G.
PY - 2007/9/3
Y1 - 2007/9/3
N2 - 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.
AB - 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.
KW - Allosteric enzymes
KW - Anti-HIV antibodies sensing
KW - Immobilized engineered β-galactosidase
U2 - https://doi.org/10.1016/j.enzmictec.2007.04.001
DO - https://doi.org/10.1016/j.enzmictec.2007.04.001
M3 - Article
SN - 0141-0229
VL - 41
SP - 492
EP - 497
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
ER -