Synthesis and application method of antigens for various heavy metals
A synthesis method and heavy metal technology, applied in the field of environmental pollutant detection, can solve the problems of few reports on antigen synthesis and identification research, difficulty in finding synthesis and identification methods, and different technical details of the methods, so as to ensure food safety and people's safety. Good health, good application prospects and significant effects
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Embodiment 1
[0021] Preparation of heavy metal cadmium complete antigen
[0022] Such as figure 1 As shown, it is the steps of this example, wherein: the amino group on the benzene ring of the bifunctional chelating agent p-NH2-Bn-DTPA and the amino group on the carrier protein are connected through glutaraldehyde to chelate heavy metals to synthesize artificial antigens. The details are as follows:
[0023] Prepare p-NH2-Bn-DTPA 10mg dissolved in 3mL HEPES buffer (10mM, pH 7.4), carrier protein BSA15mg or KLH 30mg dissolved in 6mL HEPES buffer, glutaraldehyde (GA) 50% aqueous solution with HEPES buffer 100 times Dilute, slowly add the p-NH2-Bn-DTPA solution to the carrier protein solution at room temperature, stir while dropping, then slowly add 1 mL of diluted GA into the reaction solution, stir while dropping, and stir overnight at room temperature . The next day, the reaction solution was colorless or light yellow and transparent. Use a 15mL centrifugal ultrafiltration tube (molecul...
Embodiment 2
[0026] Preparation of heavy metal lead complete antigen
[0027] After dissolving 10mg p-NH2-Bn-DTPA, add dropwise to carrier protein BSA 15mg or KLH 30mg, then add dropwise 100 times diluted 50% glutaraldehyde aqueous solution 1mL, and stir at room temperature overnight. Use a 15mL centrifugal ultrafiltration tube for ultrafiltration washing 3 times, after dilution, add 2mL of 1mg / mL heavy metal lead solution dropwise, and stir at room temperature for 5 hours. The reacted solution was ultrafiltered and washed three times with a 15 mL centrifugal ultrafiltration tube to obtain the heavy metal lead complete immunogen Pb-DTPA-KLH and the coated detection original Pb-DTPA-BSA.
Embodiment 3
[0029] Preparation of Complete Antigen of Heavy Metal Mercury
[0030] After dissolving 10mg p-NH2-Bn-DTPA, add dropwise to carrier protein BSA 15mg or OVA 10mg, then add dropwise 100 times diluted 50% glutaraldehyde aqueous solution 1mL, and stir at room temperature overnight. Use a 15mL centrifugal ultrafiltration tube for ultrafiltration washing 3 times, after dilution, add 2mL of 1mg / mL heavy metal mercury solution dropwise, and stir at room temperature for 5 hours. The reacted solution was ultrafiltered and washed three times with a 15 mL centrifugal ultrafiltration tube to obtain the heavy metal mercury complete immunogen Hg-DTPA-BSA and the coated detection original Hg-DTPA-OVA.
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