بهينه‌سازي و توسعه زيست حسگر ايمني الكتروكمي¬لومينسانس به منظور رديابي نوري هورمون بارداري

Electrochemiluminescence (ECL) is a very advanced technique for analyzing biological analytes. This technique is to completely separate the input source (electricity) and the output signal (light). The aim of this study was to develop and manufacture ECL-HCG kit components for the quantitative detection of pregnancy hormone (HCG) using an electrochemiluminescence (ECL) optical biosensor based on magnetic nanoparticles based on bipolar closed cell. In fact, this kit has two main components: iron nanoparticles coated with gold and attached to alpha HCG antibody and beta HCG antibody attached to biotin. Finally, the HCG kit was eva‎luated in the electrochemi-luminescence measuring device. From the XRD characterization results for Fe3O4, we found that it gives strong peaks at an angle of 60.35 degrees and according to standard card No. 0629-19 Fe3O4. Also, DLS and Zeta potential results for Citrate-Fe3O4 and Fe3O4 nanoparticles show the hydrodynamic diameter of 162.8 ± 39.8 nm and 169.3 ± 53.8 nm, respectively, as well as zeta potential equal to -39.7 mv and zeta potential equal to -40.6 mv. Also, the UV-VIS results for the characterization of Fe3O4-Au and Fe3O4 nanoparticles showed that Fe3O4 Au has a peak at around 580 to 600 nm, but Fe3O4 did not have any peak in this range. ICP-MS characterization results were used for the concentration of Fe3O4-Au elements and it showed that there were 87.622 ppm gold and 24.742 ppm iron in the solution. Also, the FTIR results for the characterization of Fe3O4, Fe3O4-Au, Fe3O4-Au-αHCG and biotin-βAb indicate the presence of pure iron peak and carboxylic peaks for gold coating and amide group peaks for carboxyl bonding of nanoparticles and amine group of alpha antibody and binding of carboxyl groups of biotin to N-H groups of beta antibody. Also, the results of SDS-PAGE for the characterization of Fe3O4-Au-αHCG and biotin βAb indicate the presence of antibody bound to nanoparticles and biotin in KD 73 and KD 75 bands, respectively. Finally, Bradford results for characterizing Fe3O4-Au-αHCG and biotin-βAb concentrations were 1622 µg/mL and 89.75 µg/mL, respectively. According to the methodology carried out, the kit made by us has the ability to be a diagnostic tool for HCG analytes, and based on the test, it was possible to identify and provide optical signals with measurable quality.

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