A kind of synthesis method of quantum dot-polypeptide complex with polyhistidine linking arm
A technique of polypeptide complex and polyhistidine, which is applied to the preparation method of peptides, chemical instruments and methods, peptides, etc.
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[0012] Preparation of histidine-N-endocarboxylic acid anhydride
[0013] Weighing: 3.103g histidine and 2.8g triphosgene were added into a three-neck flask. Add dry tetrahydrofuran into the three-necked flask. Stir with a magnetic stirrer and nitrogen gas slightly. The oil bath is controlled to be heated at 65° C., so that the reaction system reaches a reflux state. After about 10 to 20 minutes, the reaction solution was clear and transparent, and then nitrogen gas was blown vigorously for about half an hour. When the reaction is terminated, the liquid is clear, transparent and pale yellow. Quickly remove the oil bath. The reaction solution was poured into 250 mL of n-hexane under stirring while it was hot. Mix well and a large amount of white solid forms. The crude product was recrystallized from tetrahydrofuran and n-hexane, and it was a tiny needle-like white powder. It was filtered and vacuum-dried. The yield: 77.9%.
Embodiment 1
[0015] Synthesis of porcine enterotoxin-derived Escherichia coli K88ac adhesion antigen-8-unit histidine arm-quantum dot complex:
[0016] Mix 0.01 mol of porcine enterotoxin-derived Escherichia coli K88ac adhesion antigen and 0.08 mol of histidine-N-endocarboxylic acid anhydride in DMF solution, stir at room temperature for 1 hour, and continuously pass argon gas into the solution during this period, and pour the reaction solution Pour into n-hexane to precipitate a white colloidal solid, and vacuum dry the filter cake after filtration.
[0017] CdSe / ZnS shell quantum dots were selected and dispersed in phosphate buffer at room temperature with a quantum dot concentration of 0.1mol / L. Use N-hydroxysulfosuccinic acid imide as the activator at a concentration of 20 mol / L, react on a shaking table at room temperature for 1 hour, wash with neutral acetonitrile to remove excess activator. Antibody polypeptide chain-histidine arm: porcine enterotoxin-derived Escherichia coli K88ac...
Embodiment 2
[0019] Synthesis of hepatitis B virus core antigen-300 unit histidine arm-quantum dot complex:
[0020] Mix 0.001 mol of hepatitis B virus core antigen and 0.30 mol of histidine-N-endocarboxylic acid anhydride in water / acetonitrile solution, stir at room temperature for 2 hours, during which argon gas is continuously flowed into the solution, and the reaction solution is poured into n-hexane A white colloidal solid was precipitated out, and the filter cake was vacuum-dried after filtration.
[0021] CdSe / ZnS shell quantum dots were selected and dispersed in phosphate buffer at room temperature with a quantum dot concentration of 0.1mol / L. Use N-hydroxysulfosuccinic acid imide as the activator at a concentration of 20 mol / L, react on a shaking table at room temperature for 1 hour, wash with neutral acetonitrile to remove excess activator. Hepatitis B virus core antigen - 300 unit histidine arm. Add the above-mentioned phosphate buffer, and centrifuge the resulting reaction pr...
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