XXVII. BALKAN CLINICAL LABORATORY FEDERATION MEETING BCLF 2019 XXX. NATIONAL CONGRESS OF THE TURKISH BIOCHEMICAL SOCIETY TBS 2019, Antalya, Turkey, 27 October 2019 - 31 January 2020, vol.44, pp.67
Objectives: Gathering magnetic beads (MBs) and screen printed
electrode technology allows a highly sensitive and innovative methodology for
the development of amperometric immunosensors. These researchs purpose are the
detection of β-casomorphine-7 peptide in cheese rennet and analyze the
substrate effect on HRP-based amperometric immunosensors by using this combined
technology.
Material and Methods: Both study were based on direct and competitive immunosensor
protocols in which antigen-antibody and labeled enzyme immunoassay procedure
was followed. HRP substrates ABTS and TMB were used to measure the HRP activity
as the last step of the amperometric immunosensor analysis. Signal of the
biosensors was in nano amper (nA) range.
Results: For BCM-7 detection, competitive type immunosensor
sensitivity was found in ng/ml range and the detection limits were found as
0.5-200 ng/ml. BCM-7 detection in different commercially available cheese
rennets was also done. For HRP-based enzymatic sensor development, it is found
that TMB is more sensitive than ABTS as the substrate of the HRP.
Conclusions: BCM-7 detection in cheese rennet with MB-SPE based
immunosensor is the first study in the literature. For the other study, HRP
activity detection with immunosensor type biosensor by using different
substrates gives us the best substrate for HRP activity determinations in
amperometric detection.
Keywords: Amperometric immunosensor, BCM-7, HRP, ABTS, TMB
Acknowledgements
These
studies were supported by Erciyes University Scientific Research Projects Unit
under the code of FYL-2018-8413 and Tübitak 1509, 9130058, PrintECELISA project.