Determination of sFlt-1:angiogenic factor complex
View Patent ↗Methods for determining the presence or amount of a complex comprising a first and second molecular entity are provided, preferably an sFlt-1:PlGF complex. A determination of the presence or amount of the complex can be used in methods for predicting, detecting, monitoring a disease, or guiding therapy in respect to a disease such as vascular, vascular-related disease, cardiac, cardiac-related disease, cancer, cancer-related disease, preeclampsia, and preeclampsia-related disease. Determining sFlt-1:angiogenic factor complex is particularly useful for predicting and detecting preeclampsia in early stages of gestation and in stages of the disease where clinical evaluation may be uninformative.
1. A method of determining the presence or amount of sFlt-1:PlGF complex, the method comprising:
a) capturing the complex with a single capture agent that binds to PlGF, and detecting the complex with a single labeled detector agent that binds to one or both of sFlt-1 and sFlt-1:PlGF complex; or
b) capturing the complex with a single capture agent that binds to sFlt-1, and detecting the complex with a single labeled detector agent that binds to one or both of PlGF and sFlt-1:PlGF complex; or
c) capturing the complex with a single capture agent that binds to sFlt-1:PlGF complex, and detecting the complex with a single labeled detector agent that binds to sFlt-1:PlGF complex, wherein the single capture agent and the single labeled detector agent are capable of being bound simultaneously to the complex; or
d) capturing the complex with a single capture agent that binds to sFlt-1:PlGF complex, and detecting the complex with a single labeled detector agent that binds to one or both of PlGF and sFlt-1;
wherein the method uses a composition consisting of the single capture agent and the single labeled detector agent.
2. The method of claim 1 wherein PlGF is PlGF-1, PlGF-2, or PlGF-3.
3. The method of claim 1 wherein the single capture agent is immobilized on a support or capable of being immobilized on a support.
4. The method of claim 3 wherein the support is a surface of a test tube, a microwell, a microtiter well, a membrane, a sheet, a bead, a microparticle, or a chip.
5. The method of claim 3 wherein the single capture agent is linked to biotin.
6. The method of claim 5 wherein the single capture agent linked to biotin is immobilized through binding to avidin, streptavidin, or other biotin-binding agent.
7. The method of claim 1 wherein one or both of the single capture agent and the single labeled detector agent are polyclonal antibodies.
8. The method of claim 1 wherein one or both of the single capture agent and the single labeled detector agent are monoclonal antibodies.