反应详情

A solution of 7-mercapto-2-propylheptanoic acid(prepared in Example 2, 150 mg) in 0.1 M phosphate buffer (pH 7.0,10 ml) was added to a solution of BSA(350 mg) in 0.1 M phosphate buffer (pH 7.0,30 ml) in an argon atmosphere. To the solution was added slowly a solution of N-(m-maleimidobenzoyloxy)succinimide (hereinafter referred to as m-MBS, 150 mg) in dioxane (10 ml). The mixture was stirred for 3 hours at ambient temperature, and dialyzed overnight against running water. The dialized solution was adjusted to pH 8.5 with 1 N sodium hydroxide solution and concentrated to about 10 ml by ultrafiltration using PM30 membrane (Amicon Corp.). The concentrate was adjusted to pH 4.5 with diluted hydrochloric acid and left in a cold place for 20 hours. The resulting precipitates were collected by centrifuge (10000 r.p.m., 15 minutes), suspended in a small amount of water, dissolved by adjusting to pH 8.5 with diluted alkali solution and dialyzed for 48 hours against 0.9% sodium chloride solution (2 liters), while the outer solution was renewed four times. The dialyzed solution was lyophilized to give the conjugate (330 mg).

1. customdialyzed overnight
2. concentrationconcentrated to about 10 ml by ultrafiltration
3. waitleft in a cold place for 20 hours
4. customThe resulting precipitates
5. customwere collected by centrifuge (10000 r.p.m
6. custom15 minutes
7. dissolutiondissolved
8. additionwith diluted alkali solution
9. customdialyzed for 48 hours against 0.9% sodium chloride solution (2 liters), while the outer solution
10. customto give the conjugate (330 mg)