Reaction #2487409

ord-cace795e082b4b5dbc52f297c512b377

Reaction equation

O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
D-Glucose
OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O
D-maltose
N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
D-glucosamine
CC(=O)N[C@@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
N-acetyl-D-mannosamine
→
OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@@H](O[C@H]3[C@H](O)[C@@H](O)[C@@H](O)O[C@@H]3CO)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O
maltotriose

Reactants

O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
D-Glucose
OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O
D-maltose
N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
D-glucosamine
CC(=O)N[C@@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
N-acetyl-D-mannosamine
—
O-(3-thiolpropyl)hydroxylamine
—
solution

Reagents

None

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    OtherProgress of reaction
  2. 2
    OtherMaltotriose-SH (30 mg) was separated from linker
  3. 3
    Otherfreeze-dried
  4. 4
    OtherNext 30 mg of maltotriose-SH was reacted with bromoacetamido-derivatized BSA (15 mg)
  5. 5
    Otherprepared as above

Procedure

D-Glucose (10 mg), D-maltose (10 mg) maltotriose (25 mg), D-glucosamine (10 mg) or N-acetyl-D-mannosamine (10 mg) were reacted with O-(3-thiolpropyl)hydroxylamine (6 mg) in 1 ml D2O adjusted to pH 5.5 with 30% solution of NaOD at 37° C. for 15 hours. Progress of reaction was monitored by 1H NMR. Maltotriose-SH (30 mg) was separated from linker by passing through BioBel P-2 column in 0.05 M pyridine acetate buffer as above and freeze-dried. Next 30 mg of maltotriose-SH was reacted with bromoacetamido-derivatized BSA (15 mg), prepared as above to form maltotriose-BSA conjugate by thioether linkages. Reaction was done in buffer A, pH 7.4, 3 hours and solution was purified on Sephadex G-100 column as above. Extent of conjugation was evaluated by MALDI-TOF. Molecular mass of bromoacetamido-BSA was 73545 Da, while maltotriose-BSA conjugate was 81673 Da, indicated incorporation of 16 maltotriose molecules per BSA.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US08795680B2uspto-grants-2014_08