Kits for the diagnosis of pre-eclampsia or eclampsia

Patent metadata

Field	Value
Publication number	US-9518992-B2
Application number	US-22024508-A
Country	US
Kind code	B2
Filing date	Jul 23, 2008
Priority date	Jul 19, 2002
Publication date	Dec 13, 2016
Grant date	Dec 13, 2016

Abstract

Disclosed herein are kits for diagnosing pre-eclampsia and eclampsia or a propensity to develop pre-eclampsia or eclampsia that include agents for the detection of levels of free placental growth factor in a subject.

First claim

What is claimed is:

1. A diagnostic kit for use in diagnosing pre-eclampsia or eclampsia or a propensity to develop pre-eclampsia or eclampsia in a subject, said kit comprising:
(a) a first antibody, or antigen-binding fragment thereof, which specifically binds free placental growth factor (PlGF), wherein said first antibody, or antigen-binding fragment thereof, is detectably labeled or coupled to an enzyme;
(b) a normal reference sample for PlGF, wherein said normal reference sample for PlGF is a sample containing a level of PlGF found in a pregnant subject not having pre-eclampsia or eclampsia or a propensity to develop pre-eclampsia or eclampsia; a subject not having early-onset pre-eclampsia or eclampsia; or a subject that is not pregnant, wherein the normal reference sample for PlGF comprises purified or recombinant PlGF at a concentration of 400 to 800 pg/ml;
(c) a second antibody, or antigen-binding fragment thereof, which binds soluble Flt-1 (sFlt-1), wherein said second antibody or antigen-binding fragment thereof is detectably labeled or coupled to an enzyme;
(d) a normal reference sample for sFlt-1, wherein said normal reference for sFlt-1 sample is a sample containing a level of sFlt-1 found in a pregnant subject not having pre-eclampsia or eclampsia or a propensity to develop pre-eclampsia or eclampsia; a subject not having early onset pre-eclampsia or eclampsia; or a subject that is not pregnant, and wherein said normal reference sample comprises purified or recombinant sFlt-1 at a concentration of 0.9-2 ng/ml; and
(e) instructions for use of the kit for the diagnosis of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia in a subject, wherein said instructions comprise a step for comparison between a subject sample and the normal reference sample of (b) and (d), wherein a subject sample with a level of free PlGF that is less than the normal reference sample of step (b) and a level of sFlt-1 that is greater than the normal reference sample of step (d) is diagnostic of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia.

2. A diagnostic kit for use in diagnosing pre-eclampsia or eclampsia or a propensity to develop pre-eclampsia or eclampsia in a subject, said kit comprising:
(a) a first antibody, or antigen-binding fragment thereof, which specifically binds free placental growth factor (PlGF), wherein said first antibody or antigen-binding fragment thereof is detectably labeled or coupled to an enzyme;
(b) a positive reference sample for PlGF, wherein said positive reference sample for PlGF is a sample containing a level of PlGF found in a pregnant subject having pre-eclampsia, eclampsia, or early-onset pre-eclampsia or a propensity to develop pre-eclampsia, eclampsia, or early-onset pre-eclampsia, wherein the positive reference sample for PlGF comprises purified or recombinant PlGF at a concentration of less than 400 pg/ml;
(c) a second antibody, or antigen-binding fragment thereof, which binds soluble Flt-1 (sFlt-1), wherein said second antibody or antigen-binding fragment thereof is detectably labeled or coupled to an enzyme;
(d) a positive reference sample for sFlt-1, wherein said positive reference sample for sFlt-1 is a sample containing a level of sFlt-1 found in a pregnant subject having pre-eclampsia, eclampsia, or early-onset pre-eclampsia or a propensity to develop pre-eclampsia, eclampsia, or early-onset pre-eclampsia, and wherein said positive reference sample of comprises purified or recombinant sFlt-1 at a concentration of greater than 2 ng/ml; and
(e) instructions for use of the kit for the diagnosis of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia in a subject, wherein said instructions comprise a step for comparison between the subject sample and positive reference sample of (b) and (d), wherein a subject sample with a level of free PlGF that is less than or equal to the positive reference sample of step (b) and a level of sFlt-1 that is greater than or equal to the positive reference sample of step (d) is diagnostic of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia.

3. The kit of claim 1 , wherein said recombinant PlGF comprises the sFlt-1 or Flt-1 binding region of PlGF.

4. The kit of claim 1 or 2 , wherein said antibody or antigen-binding fragment thereof, of (a) specifically binds the sFlt-1 binding domain of PlGF.

5. The kit of claim 1 or 2 , wherein said antibody, or antigen-binding fragment thereof, of (a) or (c) is detectably labeled.

6. The kit of claim 5 , wherein said detectable label is a colorimetric label or a fluorescent label.

7. The kit of claim 1 or 2 , wherein said antibody or antigen-binding fragment thereof of (a) or (c) is coupled to an enzyme.

8. The kit of claim 7 , wherein said enzyme is horseradish peroxidase, 3-galactosidase, or alkaline phosphatase.

9. The kit of claim 1 or 2 , further comprising a second antibody, or antigen-binding fragment thereof, which specifically binds to said free PlGF.

10. The kit of claim 9 , wherein said second PlGF antibody, or antigen-binding fragment thereof, is coupled to an enzyme and the kit further comprises a preparation of a substrate for the enzyme.

11. The kit of claim 10 , wherein said enzyme is horseradish peroxidase, galactosidase, or alkaline phosphatase.

12. The kit of claim 1 , wherein said instructions comprise a step of determining the pre-eclampsia anti-angiogenic index (PAAI) of the subject and said kit further comprises a normal PAAI reference, wherein a subject sample with a PAAI that is greater than the normal PAAI reference is diagnostic of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia.

13. The kit of claim 1 , wherein said instructions comprise a step of determining the sFlt-1/PlGF of the subject and said kit further comprises a normal sFlt-1/PlGF reference value, wherein a subject sample with a sFlt-1/PlGF that is greater than the normal sFlt-1/PlGF reference is diagnostic of pre-eclampsia or eclampsia or the propensity to develop pre-eclampsia or eclampsia.

14. The kit of claim 1 or 2 , further comprising a membrane or a solid phase wherein said antibody or antigen-binding fragment thereof of (a) or (c) or both are immobilized on said membrane or said solid phase.

15. The kit of claim 1 or 2 , wherein said kit comprises components for an immunological assay, an enzymatic assay, a fluorescence polarization assay, or a colorimetric assay.

16. The kit of claim 1 or 2 , wherein said free PlGF of step (a) is an alternatively spliced isoform of PlGF.

17. The kit of claim 1 or 2 , wherein said sFlt-1 of step (c) is an isoform of sFlt-1.

18. The kit of claim 1 or 2 , wherein said sFlt-1 of step (c) is free or total sFlt-1.

19. The kit of claim 1 or 2 , wherein said sFlt-1 of step (c) is bound sFlt-1.

20. The kit of claim 1 , wherein said antibody, or antigen-binding fragment thereof, of (c) specifically binds sFlt-1.

Assignees

• Karumanchi S Ananth
• Sukhatme Vikas P
• Beth Israel Deaconess Medical Ct Inc

Inventors

• Karumanchi S Ananth
• Sukhatme Vikas P

Classifications

• G01N2800/368

  Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour · CPC title

• A61K31/522

  having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir · CPC title

• G01N33/689 Primary

  related to pregnancy or the gonads · CPC title

• G01N33/74

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

• A61K2039/505

  comprising antibodies · CPC title