申请人:WHITTAKER CORPORATION
公开号:EP0074761A1
公开(公告)日:1983-03-23
Disclosed are chromogenic or fluorogenic peptide-type compounds adapted for use in determining endotoxin in a sample by a Limulus amebocyte lysate-type assay. The compounds have the general formula:wherein R, represents hydrogen, a blocking aromatic hydrocarbon or acyl; A1 represents an L or D-amino acid selected from either lieu, Val or Leu; A2 represents Glu or Asp; A3 represents Ala or Cyst; A4 represents Arg, B represents a linkage group selected from ester and amide linkage groups; and R2 represents a chromogenic orfluorogenic group which is covalently attached to the C-carboxyl terminal of arginine through the B linkage group and which yields a chromophoric or fluorescent marker compound of the formula R2-B-H upon enzymatic hydrolysis from the remainder of the peptide-type compound by activated LAL pro-clot enzyme. Also disclosed are methods for the determination of endotoxin in a sample by contacting the sample with a pro-clotting enzyme from Limulus amebocyte lysate and one of the above chromogenic or fluorogenic peptide-type compounds.
本发明公开了一些发色性或发荧光性的肽类化合物,适用于通过纤毛虫卵母细胞裂解物类检测法确定样品中的内毒素。这些化合物具有通式:其中 R 代表氢、阻塞芳香烃或酰基;A1 代表选自 lieu、Val 或 Leu 的 L 或 D-氨基酸;A2 代表 Glu 或 Asp;A3 代表 Ala 或 Cyst;A4 代表 Arg,B 代表选自酯和酰胺连接基团的连接基团;以及 R2 代表发色或发荧光基团,该基团通过 B 连接基团共价连接到精氨酸的 C 羧基末端,并在活化的 LAL 原凝血酶从肽型化合物的剩余部分进行酶水解时产生式 R2-B-H 的发色或发荧光标记化合物。本发明还公开了测定样品中内毒素的方法,该方法是将样品与来自纤毛虫卵母细胞裂解物的促凝血酶和上述发色性或发荧光性肽类化合物之一接触。