Bicarbonate Ion → Biotin

HRID 7341

Reaction equation

HRID 7341 reaction equation
[C:1](=[O:4])(O)[O-:2].[CH2:5]1[S:9][C@@H:8]([CH2:10][CH2:11][CH2:12][CH2:13]C(NCCCCCC(ON2C(=O)C(S([O-])(=O)=O)CC2=O)=O)=O)[C@H:7]2[NH:36][C:37]([NH:39][C@@H:6]12)=[O:38].[Na+]>O>[OH:2][C:1]([CH2:13][CH2:12][CH2:11][CH2:10][C@H:8]1[C@@H:7]2[C@@H:6]([NH:39][C:37]([NH:36]2)=[O:38])[CH2:5][S:9]1)=[O:4]

Participants

4

Reactant 2

HCID 769Bicarbonate IonHCID 71571497Sulphosuccinimidyl-6-(biotinamido)hexanoate

Product 1

HCID 171548Biotin

Reagents, catalysts & solvents 1

HCID 962WaterSolvent

Procedure

About 2.0 mg of DBP is buffer exchanged with 20 mM bicarbonate at pH 9.6 three times to give a final volume of 1.0 mL. Two aliquots of sulfo-NHS-LC-biotin (Pierce) are dissolved in water to give a 1 mg/mL solution just prior to addition. The first aliquot (10 uL) is added and the reaction is maintained at room temperature for 5 minutes. The second aliquot (10 uL) is added and the reaction proceeds for an additional 4 minutes. The unreacted biotin is removed by size exclusion chromatography using SEPHADEX G-25 and 100 mM PBS, pH 7.4 as the mobile phase.

Workup

  1. customto give a final volume of 1.0 mL
  2. customto give a 1 mg/mL solution just
  3. additionto addition
  4. additionThe first aliquot (10 uL) is added
  5. additionThe second aliquot (10 uL) is added
  6. customThe unreacted biotin is removed by size exclusion chromatography

Sources & evidence

ORD record
ord-34edcaa7e3ba4173928e329654c5e7d9
Source dataset
uspto-grants-2006_08
Patent number
US07087395B1

Linked notes

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