PREECLAMPSIA BIOMARKERS AND RELATED SYSTEMS AND METHODS
Disclosed herein are methods, kits, tests, and systems for detecting, predicting, monitoring, or ruling out preeclampsia in pregnant women. Also provided herein are novel diagnostic markers, methods of data analysis, assay formats, and kits employing such markers to improve one or more characteristics of a test for identifying or ruling out preeclampsia based on biomarkers from patient samples.
1 . A method for avoiding unnecessary treatment of preeclampsia, the method comprising:
(a) contacting a biological sample that has been collected from a pregnant human female with a plurality of different probes, wherein the plurality of different probes comprises probes with specific affinity for four or more proteins selected from proteins listed in Table A and Table B;
(b) determining, based on binding of the plurality of different probes to corresponding proteins, an amount or concentration for each of the four or more proteins; and
(c) proceeding with treatment of said pregnant human in a manner that avoids unnecessary treatment of preeclampsia based at least in part on the amounts or concentrations of the four or more proteins determined in step (b).
2 - 36 . (canceled)
37 . A method for detecting and/or quantifying a plurality of proteins in a sample from a pregnant human female, the method comprising:
contacting a biological sample from a pregnant human female with a plurality of probes, wherein the plurality of probes comprises probes with specific affinity for four or more proteins wherein the four or more proteins comprise:
(a) placental growth factor;
(b) one or more angiogenesis-associated proteins selected from the group consisting of soluble fms-like tyrosine kinase 1 (sFlt1), endoglin, pappalysin 2 (PAPP-A2), and decorin; and
(c) one or more kidney damage associated-proteins selected from the group consisting of (1) kidney injury molecule-1 (KIM1), (2) programmed cell death 1 ligand 1 (CD274), and decorin; and
detecting the presence and/or quantity of the four or more proteins based on binding of the plurality of different probes to corresponding proteins.
38 . (canceled)
39 . The method of claim 38 , wherein the four or more proteins further comprise one or more proteins selected from the group consisting of C-type lectin domain family 4 member A (CLEC4A), fibroblast growth factor 21 (FGF21), trefoil factor 2 (TFF2), and hepatocyte growth factor (HGF).
40 . The method of claim 37 , wherein the four or more proteins comprise PlGF, sFlt1, KIM1, and CLEC4A.
41 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity to fibroblast growth factor 21 (FGF21), and the four or more proteins comprise FGF21.
42 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for endoglin, and the four or more proteins comprise endoglin.
43 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for decorin, and the four or more proteins comprise decorin.
44 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for cluster of differentiation 274 (CD274), and the four or more proteins comprise CD274.
45 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for hepatocyte growth factor (HGF), and the four or more proteins comprise HGF.
46 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for trefoil factor 2 (TFF2), and the four or more proteins comprises TFF2.
47 . The method of claim 37 , wherein the plurality of different probes comprises probes with specific affinity for pappalysin-2 (PAPP-A2), and the four or more proteins comprises PAPP-A2.
48 . The method of claim 37 , wherein the biological sample has been collected from the pregnant human female after gestational week 21.
49 . The method of claim 37 , wherein the biological sample has been collected from the pregnant female prior to gestational week 30.
50 . The method of claim 37 , wherein the biological sample is a urine, blood, amniotic fluid, exosome, plasma, or serum sample.
51 . The method of claim 37 , wherein the biological sample is from blood of the pregnant human female.
52 . The method of claim 37 , wherein the amount or concentration of no more than 20 proteins is determined.
53 . The method of claim 37 , wherein one or more of the plurality of different probes are antibodies or antibody fragments.
54 . The method of claim 37 , wherein each of the plurality of different probes are antibodies or antibody fragments.
55 . The method of claim 37 , wherein the plurality of probes contact the biological sample or a fraction thereof in a single reaction vessel.
56 . The method of claim 37 , wherein the plurality of probes contact the biological sample or a fraction thereof in separate reaction vessels for each protein of the four or more proteins.
57 . The method of claim 37 , further comprising a second plurality of probes, wherein the second plurality of probes comprises a probe set that is specific for binding to PlGF, a probe set that is specific for binding to sFlt1, a probe set that is specific for binding to KIM1, and a probe set that is specific for binding to CLEC4A, wherein the second plurality of probes binds to its corresponding protein at an epitope that differs from the epitope to which each of the first plurality of probes binds.
58 . The method of claim 57 , wherein coincident binding of at least one pair of the first and the second plurality of probes to the same protein molecules in the sample is detected by a luminescent oxygen channeling immunoassay (LOCI), a time-resolved fluorescence resonance energy transfer (TR-FRET) assay, an amplified luminescent proximity homogenous assay, an enzyme-linked immunosorbent assay, a proximity extension assay, or a lateral flow assay.
59 - 76 . (canceled)
77 . A mixture comprising:
a fluid sample from a pregnant female subject;
a first plurality of different probes, wherein the first plurality of different probes comprises different probes, each with specific affinity for four or more proteins selected from proteins listed in Table A or Table B.
78 . The mixture of claim 77 , wherein the four or more proteins comprise:
(a) placental growth factor (PlGF);
(b) one or more angiogenesis-associated proteins selected from the group consisting of soluble fms-like tyrosine kinase 1 (sFlt1), endoglin, pappalysin 2 (PAPP-A2), and decorin; and
(c) one or more kidney damage associated-proteins selected from the group consisting of (1) kidney injury molecule-1 (KIM1), (2) programmed cell death 1 ligand 1 (CD274), and decorin.
77 - 87 . (canceled)
88 . The mixture of claim 77 wherein the fluid sample has been collected after gestational week 20.
89 - 154 . (canceled)
155 . The method of claim 37 , wherein the biological sample is obtained from the pregnant female after gestational week 28.
156 . The method of claim 37 , wherein the biological sample is obtained from the pregnant female after gestational week 30.
157 . The method of claim 37 , further comprising: applying a classifier algorithm to an expression profile of the four or more proteins, wherein the classifier algorithm calculates an index; and
comparing the index to a reference value to determine whether to avoid the unnecessary treatment of preeclampsia.
158 . The method of claim 37 , wherein the biological sample was obtained from the pregnant female after the pregnant female has shown one or more symptoms of preeclampsia, wherein the symptoms of preeclampsia are selected from (1) high blood pressure and (2) proteinuria.