A biotin-labeled polymer nanoparticle mimic antibody and its application in bt protein ELISA
A technology of biotin-labeling and nano-particles, which is applied in the field of biotin-labeling polymer nano-particle mimic antibodies and their preparation, can solve the problems of harsh storage and reaction conditions, difficulties in the preparation of natural antibodies, and poor environmental tolerance, and achieve improved Effect of selective affinity, good specificity, and increased workload
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Embodiment 1
[0045] Example 1: Synthesis of Biotin-labeled Polymer Nanoparticle Mimetic Antibody NP1
[0046]Weigh 10% acrylamide-NITIHITDTNNK-amide, 88% isopropylacrylamide and 2% N,N-methylenebisacrylamide in a 50mL round bottom flask, add 5mL ultrapure water, Sonicate for 5 minutes to fully dissolve it. Add 10mg of sodium dodecylsulfonate to the dissolved functional monomer solution (the amount of the total substance is 0.2mmol), add twice distilled water to 20mL, continue ultrasonication for 5min to fully dissolve it, then add a magnetic stirrer, and use A rubber stopper seals the round bottom flask. Place the round bottom flask on a magnetic stirrer and stir at a speed of 50 r / min. Insert a long needle connected to the nitrogen cylinder to the bottom of the solution through the rubber stopper, and insert the other short needle into the rubber stopper and place it on the top of the round-bottomed flask, and continue nitrogen gas for 30 minutes to completely remove the dissolved oxyge...
Embodiment 2
[0047] Example 2: Synthesis of biotin-labeled polymer nanoparticle mimic antibody NP2
[0048] Weigh 10% of acrylamide-NITIHITDTNNK-amide, 2% of N-(3-methacrylamidopropyl)-D-biotinamine, 86% of isopropylacrylamide and 2% of Add 8mL of ultrapure water to a 50mL round-bottomed flask of N,N-methylenebisacrylamide, and sonicate it for 8min to fully dissolve it. Add 10 mg of sodium dodecylsulfonate to the dissolved functional monomer solution (the amount of the total substance is 0.2 mmol), add twice distilled water to 20 mL, continue ultrasonication for 3 min to fully dissolve, then add a magnetic stirrer, and use A rubber stopper seals the round bottom flask. Place the round bottom flask on a magnetic stirrer and stir at a speed of 160r / min. Insert a long needle connected to the nitrogen cylinder to the bottom of the solution through the rubber stopper, and insert the other short needle into the rubber stopper and place it on the top of the round-bottomed flask, and continue ni...
Embodiment 3
[0049] Example 3: Synthesis of biotin-labeled polymer nanoparticle mimic antibody NP3
[0050] Weigh 10% of acrylamide-NITIHITDTNNK-amide, 4% of N-(3-methacrylamidopropyl)-D-biotinamine, 84% of isopropylacrylamide and 2% of Add 5mL of ultrapure water to a 50mL round-bottomed flask of N,N-methylenebisacrylamide, and sonicate for 5min to fully dissolve it. Add 10mg of sodium dodecylsulfonate to the dissolved functional monomer solution (the amount of the total substance is 0.2mmol), add twice distilled water to 20mL, continue ultrasonication for 5min to fully dissolve it, then add a magnetic stirrer, and use A rubber stopper seals the round bottom flask. Place the round bottom flask on a magnetic stirrer and stir at a speed of 300r / min. Insert a long needle connected to the nitrogen cylinder to the bottom of the solution through the rubber stopper, and insert the other short needle into the rubber stopper and place it on the top of the round-bottomed flask, and continue nitrog...
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