Piezometric biosensors for anti-apoptotic protein survivin based on buried positive-potential barrier and immobilized monoclonal antibodies.
The anti-apoptotic protein survivin (Sur) plays an important role in the regulation of cell division and inducing the chemotherapeutic drug resistance. The Sur protein and its mRNA have recently been studied as cancer biomarkers and potential targets for cancer therapy. In this work, we have focused on the design of immunosensors for the detection of Sur based on buried positive-potential barrier layer structure and anti-survivin antibody. The modification of solid AuQC piezoelectrodes was monitored by recording the resonance frequency shift and electrochemical measurements during each step of the sensor preparation. Our results indicate that the immunosensor with covalently bound monoclonal anti-survivin antibody can detect Sur with the limit of detection, LOD=1.7nM (S/N=3σ). The immunosensor applicability for the analysis of real samples was assessed by testing samples of cell lysate solutions obtained from human astrocytoma (glioblastoma) U-87MG cell line, with the experiments performed using the standard addition method. The good linearity of the calibration curves for PBS and lysate solutions at low Sur concentrations confirm the high specificity of the proposed biosensor and good discrimination against nonspecific interactions with lysate components. The calculations indicate that there is still room to increase the Sur capture capacity for Sur while miniaturizing the sensor. The important advantage of the sensor is that it can be reused by a simple regeneration procedure.
Department of Biophysics, Warsaw University of Life Sciences, SGGW, 02-776 Warsaw, Poland. Electronic address: magdalena_stobiecka@sggw.pl.
Magdalena
M
Stobiecka
Institute of Nanoparticle Nanocarriers, 11010 Barczewo, Poland.
Agata
A
Chalupa
Department of Biophysics, Warsaw University of Life Sciences, SGGW, 02-776 Warsaw, Poland.
Beata
B
Dworakowska
D055910:Antibodies, Immobilized / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation*; D045744:Cell Line, Tumor; D055664:Electrochemical Techniques / Q000295:instrumentation; D004867:Equipment Design; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D051034:Inhibitor of Apoptosis Proteins / Q000032:analysis*; D057230:Limit of Detection; D010084:Oxidation-Reduction; D058525:Quartz Crystal Microbalance Techniques / Q000295:instrumentation*; D000077022:Survivin
D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D000077022:Survivin
Immunosensor; Monoclonal anti-survivin antibody; Piezoimmunosensor; Protein survivin; Quartz crystal microbalance