反应详情

Amine functionalized polymer of formula (Vc) is dissolved at 10 mg/mL in anhydrous DMF and divided into two portions. NHS-biotin (0.9 mg in 90 μL, 88 equivalents) (Pierce, 20217) and NHS-LC-LC-biotin (Pierce, 21343) at 10 mg/mL (1.5 mg in 150 μL, 88 equivalents) are dissolved in anhydrous DMF. The NHS-biotin and NHS-LC-LC-biotin solutions are immediately added to the two portions of polymer solution and allowed to mix on a shaker overnight in the dark. Excess reactant is removed by washing the solution using Amicon Ultra-4 mL 10 kD MWCO filter cartridges in a series of wash steps: First, the cartridge is first filled approximately halfway with water, and the reaction solution (by pipet) subsequently added directly into the water. Next, the cartridge is filled with water until it is full. The solution is mixed by pipetting up and down. Then, the cartridge is centrifuged at 2400 rpm for 30 minutes, or until the volume is reduced to 250 μL. The cartridge is then refilled with water 1 drop of 1M HCl is added; the solution is mixed, and centrifuged at 2400 rpm for 30 minutes, or until the volume was reduced to 250 μL. Next, the cartridge is refilled with water, 1 drop of 1M NaOH is added; the solution is mixed, and centrifuged at 2400 rpm for 30 minutes, or until the volume is reduced to 250 μL. The cartridge is then refilled with water, mixed and centrifuged at 2400 rpm for 30 minutes, or until the volume is reduced to 250 μL. This final step is repeated for a total of 5 washes.

1. additionto mix on a shaker overnight in the dark
2. customExcess reactant is removed
3. washby washing the solution
4. filtrationfilter cartridges in a series of wash steps
5. additionFirst, the cartridge is first filled approximately halfway with water
6. additionthe reaction solution (by pipet) subsequently added directly into the water
7. additionNext, the cartridge is filled with water until it
8. additionThe solution is mixed
9. additionThe cartridge is then refilled with water 1 drop of 1M HCl
10. additionis added
11. additionthe solution is mixed
12. waitcentrifuged at 2400 rpm for 30 minutes
13. additionNext, the cartridge is refilled with water, 1 drop of 1M NaOH
14. additionis added
15. additionthe solution is mixed
16. waitcentrifuged at 2400 rpm for 30 minutes
17. additionThe cartridge is then refilled with water
18. additionmixed and centrifuged at 2400 rpm for 30 minutes