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
These studies were supported by Erciyes University Scientific Research Projects Unit under the code of FYL-2018-8413 and Tübitak 1509, 9130058, PrintECELISA project.