Method for the determination of the formation of endothelins for medical diagnostic purposes, and antibodies and kits for carrying out such a method
Described herein is an in vitro method for the determination of the formation of endothelins in serious diseases, in particular cardiovascular diseases, inflammations, sepsis and cancer, in whole blood, plasma or serum of a human patient, in which relatively long-lived peptide fragments of the processed primary prepro- or proendothelins are determined which contain neither the actual biologically active endothelin nor its direct precursor big endothelin. In particular, disclosed are antibodies and kits for selectively detecting these fragments.
1. An isolated antibody which binds specifically to a peptide consisting of amino acids (i) 184-203 or (ii) 200-212 of preproendothelin-1 (SEQ ID NO: 1).
2. The antibody of claim 1 , wherein said antibody is affinity-purified using (i) peptide 184-203 or (ii) peptide 200-212, respectively, of preproendothelin-1 (SEQ ID NO: 1).
3. A kit for, detecting C-terminal fragments of amino acids 168-212 of preproendothelin-1 (SEQ ID NO: 1), said kit comprising
(a) a first antibody selected from antibodies that specifically bind to (i) a peptide consisting of amino acids 168-181 or (ii) antibodies that specifically bind to a peptide consisting of amino acids 184-203 of preproendothelin-1 (SEQ ID NO: 1); and
(b) a second antibody selected from antibodies that specifically bind to a peptide consisting of amino acids 184-203 or antibodies that specifically bind to a peptide consisting of amino acids 200-212 of preproendothelin-1 (SEQ ID NO: 1), wherein said second antibody is different from said first antibody.
4. The kit of claim 3 , wherein one of said first and second antibodies is immobilized on the walls of a test tube.
5. The kit of claim 3 , wherein at least one of said first and second antibodies has a detectable marker and the other is immobilized or immobilizable on a solid support.
6. The kit of claim 3 , further comprising (c) a standard peptide comprising at least amino acids 168-203 or 168-212 of preproendothelin-1 (SEQ ID NO: 1).
7. The isolated antibody of claim 1 , wherein said antibody specifically binds to a peptide consisting of amino acids 184-203 of preproendothelin-1 (SEQ ID NO: 1).
8. The isolated antibody of claim 1 , wherein said antibody specifically binds to a peptide consisting of amino acids 200-212 of preproendothelin-1 (SEQ ID NO: 1).
9. The kit of claim 3 , comprising
(a) a first antibody that specifically binds to a peptide consisting of amino acids 168-181; and
(b) a second antibody that specifically binds to a peptide consisting of amino acids 184-203 of preproendothelin-1 (SEQ ID NO: 1).
10. The kit of claim 9 , further comprising (c) a standard peptide comprising at amino acids 168-203 of preproendothelin-1 (SEQ ID NO: 1).
11. The kit of claim 3 , comprising
(a) a first antibody that specifically binds to a peptide consisting of amino acids 168-181; and
(b) a second antibody that specifically binds to a peptide consisting of amino acids 200-212 of preproendothelin-1 (SEQ ID NO: 1).
12. The kit of claim 11 , further comprising (c) a standard peptide comprising at least amino acids 168-212 of preproendothelin-1 (SEQ ID NO: 1).
13. The kit of claim 3 , comprising
(a) a first antibody that specifically binds to a peptide consisting of amino acids 184-203; and
(b) a second antibody that specifically binds to a peptide consisting of amino acids 200-212 of preproendothelin-1 (SEQ ID NO: 1).
14. The kit of claim 13 , further comprising (c) a standard peptide comprising at least amino acids 168-212 of preproendothelin-1 (SEQ ID NO: 1).