m-Maleimidobenzoyl-N-hydroxysuccinimide → 7-Mercapto-2-propylheptanoic acid

HRID 325819ノート

反応式

HRID 325819 の反応式
C([S:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH:10]([CH2:14][CH2:15][CH3:16])[C:11]([OH:13])=[O:12])(=O)C.C1(=O)N(C2C=C(C=CC=2)C(ON2C(=O)CCC2=O)=O)C(=O)C=C1>P([O-])([O-])([O-])=O.O1CCOCC1>[SH:4][CH2:5][CH2:6][CH2:7][CH2:8][CH2:9][CH:10]([CH2:14][CH2:15][CH3:16])[C:11]([OH:13])=[O:12]

参加物

5

反応物 2

HCID 93861m-Maleimidobenzoyl-N-hydroxysuccinimide2 回HCID 204628707-Acetylthio-2-propylheptanoic acid

生成物 1

HCID 204628737-Mercapto-2-propylheptanoic acid

試薬・触媒・溶媒 2

HCID 1061Phosphate溶媒2 回HCID 312751,4-Dioxane溶媒

実験手順

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).

後処理

  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)

出典と根拠

ORDレコード
ord-1743572c25c349678afbf3a6ef00f8e1
出典データセット
uspto-grants-1984_04
特許番号
US04443365

反応ノート

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