N-Hydroxysuccinimido-d-biotin → Biotin

HRID 1606890Notes

Reaction equation

HRID 1606890 reaction equation
O[N:2]1[C:23](=[O:24])[NH:22][C@H:21]2[C@@H:3]1[CH2:4][S:5][C@H:6]2[CH2:7][CH2:8][CH2:9][CH:10](N1C(=O)CCC1=O)[C:11](=[O:13])[OH:12]>OP([O-])(O)=O.OP([O-])([O-])=O.[Na+].[Na+].[Na+].[Cl-].[Cl-].[K+].[K+].CN(C)C=O>[OH:13][C:11]([CH2:10][CH2:9][CH2:8][CH2:7][C@H:6]1[C@@H:21]2[C@@H:3]([NH:2][C:23]([NH:22]2)=[O:24])[CH2:4][S:5]1)=[O:12]

Participants

4

Reactant 1

HCID 129676281N-Hydroxysuccinimido-d-biotin2×

Product 1

HCID 171548Biotin

Reagents, catalysts & solvents 2

HCID 6228DimethylformamideSolventHCID 24978514Phosphate Buffered SalineSolvent

Procedure

Synthesis of biotinylated rabbit IgG was carried out as follows: rabbit IgG (2 mg, 12.5 nmoles) was dissolved in 2 mls of pH=7.4 phosphate buffered saline. N-hydroxysuccinimidobiotin was dissolved in N,N-dimethylformamide and 0.3 mg (880 nmoles) of N-hydroxysuccinimidobiotin was added and incubated at room temperature for two hours. The biotinylation mixture was purified by chromotography on Sephadex G-25 (1×25 cm), using the same buffer as eluent resulting in separation of biotinylated antibody from free biotin. The biotin: antibody ratio was 16, as determined by measuring antibody concentration by absorbance at 280 nm and measuring biotin concentration by the method of Green, N. M., Methods in Enzymology, McCormack D. B. and L. B. Wright (ed.) 18A:418 (1970).

Workup

  1. customSynthesis of biotinylated rabbit IgG
  2. customThe biotinylation mixture was purified by chromotography on Sephadex

Sources & evidence

ORD record
ord-a740a2780de94378bb5b42a537dab9d3
Source dataset
uspto-grants-1989_09
Patent number
US04863876

Reaction notes

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