NHS-Biotin → Biotin

HRID 1254967

反応式

HRID 1254967 の反応式
C1C(=O)N([O:8][C:9]([CH2:11][CH2:12][CH2:13][CH2:14][CH:15]2[S:19][CH2:18][CH:17]3[NH:20][C:21]([NH:23][CH:16]23)=[O:22])=[O:10])C(=O)C1>CN(C=O)C>[OH:10][C:9]([CH2:11][CH2:12][CH2:13][CH2:14][C@H:15]1[C@@H:16]2[C@@H:17]([NH:20][C:21]([NH:23]2)=[O:22])[CH2:18][S:19]1)=[O:8]

参加物

3

反応物 1

HCID 434213NHS-Biotin2 回

生成物 1

HCID 171548Biotin

試薬・触媒・溶媒 1

HCID 6228Dimethylformamide溶媒2 回

実験手順

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.

後処理

  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

出典と根拠

ORDレコード
ord-1919097fbb9b40e898be994a052d6f47
出典データセット
uspto-grants-2013_06
特許番号
US08455613B2

関連ノート

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