Borate → Biotin

HRID 1031062Notes

Reaction equation

HRID 1031062 reaction equation
C1C(=O)N(OC(CCCCCN[C:17]([CH2:19][CH2:20][CH2:21][CH2:22][CH:23]2[S:27][CH2:26][CH:25]3[NH:28][C:29]([NH:31][CH:24]23)=[O:30])=[O:18])=O)C(=O)C1.B([O-])([O-])[O-:33]>[Na+].[Cl-]>[OH:33][C:17]([CH2:19][CH2:20][CH2:21][CH2:22][C@H:23]1[C@@H:24]2[C@@H:25]([NH:28][C:29]([NH:31]2)=[O:30])[CH2:26][S:27]1)=[O:18]

Participants

4

Reactant 2

HCID 26574BorateHCID 5063989Biotin-X-NHS

Product 1

HCID 171548Biotin

Reagents, catalysts & solvents 1

HCID 5234Sodium ChlorideSolvent

Procedure

A suspension of XO in 3.2M (NH4)2SO4 (20 mg/ml) (Boehringer Mannheim) was centrifuged at 5000g during 10 minutes and the pellet was dissolved in 1 ml pH 8.5 50 mM, NaCl, 20 mM borate buffer. A solution of biotin X NHS (Biotin ε-aminocaproic acid N hydroxysuccinimide ester, Calbiochem) (1.2 mg/12 12 μl anhydrous dimethylformamide) was added. The solution was agitated by vortexing for two minutes, left at ambient temperature for one hour, and then dialyzed against 0.13M NaCl, 0.07M Na phosphate buffer pH 7.4 (Phosphate buffered saline PBS) containing 0.1% Na azide. The XO-biotin conjugate was then kept at +4° C. under sterile conditions.

Workup

  1. waitleft at ambient temperature for one hour
  2. customdialyzed against 0.13M NaCl, 0.07M Na phosphate buffer pH 7.4 (Phosphate buffered saline PBS)
  3. additioncontaining 0.1% Na azide
  4. customwas then kept at +4° C. under sterile conditions

Sources & evidence

ORD record
ord-c0f8ae7defa647f885af7055eb807c02
Source dataset
uspto-grants-1992_04
Patent number
US05108893

Reaction notes

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