BNP (1-32) epitope and antibodies directed against said epitope

Patent metadata

Field	Value
Publication number	US-10495649-B2
Application number	US-201615288005-A
Country	US
Kind code	B2
Filing date	Oct 7, 2016
Priority date	Aug 3, 2007
Publication date	Dec 3, 2019
Grant date	Dec 3, 2019

Abstract

The present invention relates to a polypeptide carrying a human BNP(1-32) epitope according to Formula (I): a1-R1—X1—FGRKMDR—X2—R2-a2 as well as ligands specific of the FGRKMDR epitope.

First claim

The invention claimed is:

1. A multiepitopic calibrator for the use of detecting human BNP (1-32) or human proBNP (1-108) as well as the respective fragments thereof, having the following general formula (III):
t 1 -E 1 -L 1 -E 2 [-L k-1 -E k ] n -t 2   (III)

wherein:
n is an integer between 0 and 8;
k is an integer between 3 and n+2 when n>0;
E 1 , E 2 , and E k are different from one another, wherein one of E 1 , E 2 and E k is a peptide having a sequence of R 1 —X 1 —FGRKMDR(SEQ ID NO: 8)-X 2 —R 2 , wherein
X 1 is absent or present and when present is selected among C and GC;
X 2 is absent or present and when present is selected among I and IS;
R 1 and R 2 , which may be the same or different, present or absent, represent any amino acid or a peptide chain of 2 to 15 amino acids, provided that said polypeptide of formula (III) does not include any portion of human BNP(1-32) of more than 11 amino acids including the sequence GCFGRKMDRIS (SEQ ID NO: 63),

and each of the remaining two from E 1 , E 2 and E k independently represents a sequence of 3 to 15 amino acids selected from the sequence of human proBNP(1-108) (SEQ ID NO: 1);
t 1 represents a hydrogen atom, an acetyl group, a peptide sequence of 1 to 10 amino acids, a peptide sequence of 1 to 10 N-α acetylated amino acids, a biotinyl or biocytinyl group, a peptide sequence of 1 to 10 amino acids carrying a biotinyl or biocytinyl radical, or a linear amino alkyl (C 1 -C 10 ) carbonyl chain;
t 2 represents a hydroxyl radical, an amino radical, a peptide sequence of 1 to 10 amino acids, a peptide sequence of 1 to 10 amino acids carrying a terminal amino group, or a linear or branched amino alkyl (C 1 -C 10 ) carbonyl chain; and
L 1 and L k-1 which may be the same or different, each represents a coupling agent that covalently couples the adjacent peptide chains.

2. The multiepitopic calibrator according to claim 1 , corresponding to the following general formula (IV):
t 1 -E 1 -L 1 -E 2 -t 2   (V)

wherein E 1 , E 2 , L 1 , t 1 and t 2 are as defined in claim 1 .

3. The multiepitopic calibrator according to claim 1 , wherein L 1 and L 2 represent:
—NH—(CH 2 ) 5 —CO—.

4. The multiepitopic calibrator according to claim 3 , selected from the group consisting of the multiepitopic calibrators defined by the following formulae:
Ac—YTLRAPRSPKMV(SEQ ID NO: 66)-L 1 -SFGRKMDRISS(SEQ ID NO: 64)-NH 2 ;
Ac—YTLRAPRSPKMV(SEQ ID NO: 66)-L 1 -CFGRKMDRISSSSGLGCK(SEQ ID NO: 65)-NH 2 ;
Ac—YTLRAPRSPKMVQG(SEQ ID NO: 67)-L 1 -FGRKMDR(SEQ ID NO: 8)-NH 2 ;
Ac—FGRKMDR(SEQ ID NO: 8)-L 1 -SGLGC*KVLRRH(SEQ ID NO: 68)-OH;
Ac—FGRKMDR(SEQ ID NO: 8)-L 1 -SGLGC*KVLR(SEQ ID NO: 69)-NH 2 ;
Ac-SPKMVQGSG(SEQ ID NO: 59)-L 1 -FGRKMDR(SEQ ID NO: 8)-NH 2 ;
Ac—YTLRAPRSPKMV(SEQ ID NO: 66)-L 1 -FGRKMDR(SEQ ID NO: 8)-L 2 -SGLGC*KVLRRH(SEQ ID NO: 68)-OH;
and Ac—YTLRAPRSPKMV(SEQ ID NO: 66)-L 1 -FGRKMDR(SEQ ID NO: 8)-L 2 -SGLGC*KVLR(SEQ ID NO: 69)-NH 2 ;
wherein Ac represents an acetyl group and C* represents an acetamidomethyl-blocked cysteine.

5. A kit for detecting human BNP(1-32) or human proBNP(1-108) as well as the respective fragments thereof comprising the sequence FGRKMDR (SEQ ID NO: 8), comprising at least:
a multiepitopic calibrator having the following general formula (III):
t 1 -E 1 -L 1 -E 2 [-L k-1 E k ] n -t 2   (III)

wherein:
n is an integer between 0 and 8;
k is an integer between 3 and n+2 when n>0;
E 1 , E 2 and E k are different from one another, wherein one of E 1 , E 2 and E k is a peptide having a sequence of R 1 —X 1 —FGRKMDR (SEQ ID NO: 8)-X 2 —R 2 peptide sequence, wherein
X 1 is absent or present and when present is selected among C and GC;
X 2 is absent or present and when present is selected among I and IS;
R 1 and R 2 , which may be the same or different, present or absent, represent any amino acid or a peptide chain of 2 to 15 amino acids, provided that said polypeptide of foiniula (III) does not include any portion of human BNP(1-32) of more than 11 amino acids including the sequence GCFGRKMDRIS (SEQ ID NO: 63),

and each of the remaining two from E 1 , E 2 and E k independently represents a sequence of 3 to 15 amino acids selected from the sequence of human proBNP(1-108);
t 1 represents a hydrogen atom, an acetyl group, a peptide sequence of 1 to 10 amino acids, a peptide sequence of I to 10 N-a acetylated amino acids, a biotinyl or biocytinyl group, a peptide sequence of 1 to 10 amino acids carrying a biotinyl or biocytinyl radical, or a linear amino alkyl (C 1 -C 10 ) carbonyl chain;
t 2 represents a hydroxyl radical, an amino radical, a peptide sequence of 1 to 10 amino acids, a peptide sequence of 1 to 10 amino acids carrying a terminal amino group, or a linear or branched amino alkyl (C 1 -C 10 ) carbonyl chain;
L 1 and L k-1 which may be the same or different, each represents a coupling agent that covalently couples the adjacent peptide chains.

Assignees

• Bio Rad Europe Gmbh
• Centre Nat Rech Scient

Inventors

• Rieunier Francois
• Giuliani Isabelle
• Villard-Saussine Sylvie

Classifications

• G01N2333/58

  Atrial natriuretic factor complex; Atriopeptin; Atrial natriuretic peptide [ANP]; Brain natriuretic peptide [BNP, proBNP]; Cardionatrin; Cardiodilatin · CPC title

• G01N2800/50

  Determining the risk of developing a disease · CPC title

• C07K14/58

  Atrial natriuretic factor complex; Atriopeptin; Atrial natriuretic peptide [ANP]; Cardionatrin; Cardiodilatin · CPC title

• G01N33/74 Primary

  involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors} · CPC title

• G01N2800/32

  Cardiovascular disorders · CPC title