Biomimetic Synthesis Of Nanomaterials And Their Application In ELISA For The Detection Of Bt Protein

With the rapid development of transgenic technology,genetically modified products have come into people’s sight.First of all,the safety of technology has attracted wide attention,and the supervision and safety evaluation of the genetically modified products become particularly important.Therefore,it has important practical significance for the efficient detection of genetically modified products.Biomimetic nanomaterials are a kind of organic-inorganic hybrid nanomaterials which are self-assembled by organic compounds.The synthesis method is simple and the reaction conditions are mild.At the same time,the material synthesis and enzyme immobilization are completed in one step.Because of its special synthesis process and material properties,this material has been widely used in enzyme loading.In this paper,two kinds of organic-inorganic hybrid nanomaterials were synthesized by biomimetic synthesis method,and the Btillus thuringiensis(Bt)protein Cry1 Ab was detected by enzyme-linked immunosorbent assay(ELISA).The contents of this paper are as follows:(1)The hollow bundle-like Enzyme-Brochantite(EB)organic-inorganic hybrid nanomaterials were prepared by biomimetic mineralization method.The morphology and composition of the materials were characterized by scanning electron microscopy(SEM),transmission electron microscopy(TEM),and X-ray diffraction(XRD).Then,through optimization,the optimal reaction conditions were obtained as follows: the dosage of Streptavidin labeled horseradish peroxidase(SA-HRP)was 10 μg,the dosage of biotin conjugated secondary antibody(BCSA)was 5 μg,the dilution ratio of material was 15 times,the p H value of buffer solution was 7.0,and the incubation time was 20 min.The results showed that the method had good specificity,the linear range was 0-12.5 ng/m L,the detection limit was 0.11 ng/m L.(2)The biomimetic synthesis enzyme was immobilized into the metal organic framework ZIF-8 to prepare SA-HRP@ZIF-8 Nanocomposites.The morphology and composition of the materials were characterized by scanning electron microscopy(SEM),transmission electron microscopy(TEM),and X-ray diffraction(XRD).The optimal reaction conditions were as follows: the dosage of Sstreptavidin labeled horseradish peroxidase(SA-HRP)was 20μg,the dilution ratio of BCSA was 10000 times,the dilution ratio of material was 10 times,the p H value of buffer solution was 7.0,and the incubation time was 30 min.The results showed that the method had good specificity,the linear range was 0-6.25 ng/mL,and the detection limit was 0.10 ng/mL.