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Sensitive detection of specific antibodies by biosensors has become of major importance for monitoring and controlling epidemics. Here we report a development of a biosensor able to specifically measure antibodies in a drop of unmodified blood serum. Within minutes, the detection system measures presence of antibodies against Plasmodium vivax, a causing agent for malaria. The biosensor consists of a layer of carbon nanotubes (CNTs) which were casted on a carbon working electrode area of a three-electrode system and oxidized. An amine layer was produced next by modifying the surface with EDAC/NHS followed by reaction with a diamine compound. Finally, the protein fragments derived from P. vivax containing well-known antigen sequences were casted on this layer and bound through electrostatic interactions, involving hydrogen and ionic bonding. All these chemical changes occurring at the carbon surface along the biosensor assembly were followed and confirmed by Fourier Transformed Infrared s pectrometry (FTIR) and Raman spectroscopy. The presence of antibodies in serum was detected by monitoring the electrical properties of the layer, making use of cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV), against a standard iron probe. Overall, the charge-transfer resistance decreased after antibody binding, because there was an additional amount of protein bound to the surface. This hindered the access of the iron redox probe to the conductive support at the electrode surface. Electrical changes could be measured at antibody concentration as low as ~6-50pg/L (concentrations in the range of 10-15M) and as high as ~70μg/L. Specific measurement with low background was even possible in undiluted serum. Hence, this novel biosensor allows assessing serum antibody levels in real time and in un-manipulated serum samples on-site where needed.

Original publication

DOI

10.1016/j.bios.2017.01.031

Type

Journal article

Journal

Biosensors & bioelectronics

Publication Date

05/2017

Volume

91

Pages

833 - 841

Addresses

BioMark/CINTESIS-ISEP, School of Engineering of the Polytechnique School of Porto, Portugal.

Keywords

Humans, Plasmodium vivax, Malaria, Vivax, Nanotubes, Carbon, Antibodies, Protozoan, Equipment Design, Biosensing Techniques, Electrodes, Electrochemical Techniques, Limit of Detection, HEK293 Cells, Dielectric Spectroscopy