反応 #2487409

反応方程式

D-Glucose

D-maltose

D-glucosamine

N-acetyl-D-mannosamine

→

maltotriose

反応条件

後処理

1. 1
   その他Progress of reaction
2. 2
   その他Maltotriose-SH (30 mg) was separated from linker
3. 3
   その他freeze-dried
4. 4
   その他Next 30 mg of maltotriose-SH was reacted with bromoacetamido-derivatized BSA (15 mg)
5. 5
   その他prepared as above

実験手順

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.

出典