1,2-Dichloroethane → Biotin

HRID 1175643Notes

Reaction equation

HRID 1175643 reaction equation
[CH2:1]1[S:5][C@@H:4]([CH2:6][CH2:7][CH2:8][CH2:9][C:10](NCCCCCC(NN)=O)=[O:11])[C@H:3]2[NH:22][C:23]([NH:25][C@@H:2]12)=[O:24].C(Cl)CCl.C1N(CCS(O)(=O)=O)CC[O:32]C1>CS(C)=O>[OH:32][C:10]([CH2:9][CH2:8][CH2:7][CH2:6][C@H:4]1[C@@H:3]2[C@@H:2]([NH:25][C:23]([NH:22]2)=[O:24])[CH2:1][S:5]1)=[O:11]

Participants

5

Reactant 3

HCID 111,2-DichloroethaneHCID 781654-Morpholineethanesulfonic acid2×HCID 83900N-(6-Hydrazinylidene-6-hydroxyhexyl)-5-(2-hydroxy-3a,4,6,6a-tetrahydro-1H-thieno[3,4-d]imidazol-6-yl)pentanimidic acid

Product 1

HCID 171548Biotin

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

Natural GAGs (NAH, HS, HA, HEP, Ch, CS-A(S), CS-A(W), CS-B, CS-C, CS-D, CS-E, and KS) were dissolved in 0.1 M MES buffer (pH 5.5) to prepare 10 mg/ml GAG solutions. 25 μl of 20 mM biotin-LC-hydrazide (manufactured by Pierce Biotechnology) dissolved in dimethylsulfoxide (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 1 ml of each GAG solution. Subsequently, 12.5 μl of a 100 mg/ml EDC (manufactured by Pierce Biotechnology) solution prepared with 0.1 M MES buffer (pH 5.5) was added thereto. The mixture was mixed well, and then allowed to react with stirring at normal temperature (15° C. to 25° C.) for 16 to 24 hours. After completion of the reaction, the reaction products were dialyzed by using a dialysis membrane (cutoff molecular weight: 10,000 or less) against phosphate buffered saline (pH 7.2 to 7.5, containing no divalent ions such as calcium ion: hereinafter, referred to as “PBS(−)”) as dialysis solution, to thereby sufficiently remove free biotin. After completion of dialysis, the concentration of Bi-GAG was adjusted to 5 mg/ml, followed by freeze preservation.

Workup

  1. customto prepare 10 mg/ml GAG solutions
  2. additionwas added to 1 ml of each GAG solution
  3. additionwas added
  4. additionThe mixture was mixed well
  5. customto react
  6. customAfter completion of the reaction
  7. customthe reaction products were dialyzed
  8. additioncontaining no divalent ions such as calcium ion
  9. customas dialysis solution
  10. customAfter completion of dialysis

Sources & evidence

ORD record
ord-e38685b3b95147df805acaeb5853a967
Source dataset
uspto-grants-2010_11
Patent number
US07842500B2

Reaction notes

Loading…