Determination of the amount of hLHB core fragment in a sample from a subject and uses thereof

Abstract

The present invention provides a method for predicting the likely timing of the onset of menopause for a perimenopausal female subject by determining the amount of hLHβcf in a sample from the subject comprising the steps of: (a) contacting a sample from the subject with an antibody which specifically binds to hLHβcf without substantially cross-reacting with hLH, hLHβ or hLHβcf, under conditions permitting formation of a complex between the antibody and hLHβcf; (b) measuring the amount of complex formed, so as to thereby determine the amount of hLHβcf in the sample; and (c) comprising the amount of hLHβcf in the subject's sample determined in step (b) with either (i) the amount determined for known postmenopausal female subject or (ii) the amount determined for a sample from a known premenopausal female subject, wherein an amount of hLHβcf in the sample similar to the amount of hLHβcf in the known postmenopausal sample indicates temporal proximity to the onset of menopause, and an amount of hLHβcf in the sample similar to the amount of hLHβcf in the known premenopausal sample indicates temporal distance from the onset of menopause for the subject. AS described herein amount is both concentration and pattern of measurement of concentrations in one or more consecutive urine specimens.

1 - 23 . (canceled) 
     
     
         24 . A method for assessing ovarian function in a subject comprising the steps of:
 a. contacting a sample from a subject with an antibody which specifically binds to hLHβcf without substantially cross-reacting with hLH, hLHβ or hCGβcf, under conditions permitting formation of a complex between the antibody and hLHβcf;   b. measuring the amount of complex formed, so as to thereby determine the amount of molecule in the sample; and   c. comparing the amount of hLHβcf in the subject's sample determined in step (b) with either (i) the amount determined for a sample from a subject with normal ovarian function or (ii) the amount determined for a sample from a subject with abnormal ovarian function, wherein an amount of hLHβcf in the sample similar to amount of hLHβcf in the sample from subjects having normal ovarian function indicates normal ovarian function, and amounts of hLHβcf in the sample similar to amounts of hLHβcf having abnormal ovarian function indicates abnormal ovarian function for the subject.   
     
     
         25 . The method of  claim 24 , wherein step (a) the capturing antibody specifically binds a region of hLHβcf comprising a protein portion or both a protein and a carbohydrate moiety. 
     
     
         26 . The method of  claim 24 , wherein in step (a) the antibody is bound to a solid support and in step (b) the amount of the antibody bound to the solid support in the complex with hlHβcf is measured by contacting the complex with a second antibody which binds to the complex and which is labeled with a detectable marker. 
     
     
         27 . The method of  claim 24 , wherein the sample is a urine sample, a first morning void urine sample, an aggregate sample of the first morning void urine samples for at least two consecutive days or an aggregate sample of the first morning void urine samples for five or more consecutive days, or a collection of all urinations consecutively with time noted for 24-48 hours. 
     
     
         28 . The method according to  claim 24 , wherein the abnormal ovarian function is hyperactivity. 
     
     
         29 . The method according to  claim 24 , wherein the abnormal ovarian function is hypoactivity. 
     
     
         30 . The method of  claim 24 , wherein the second antibody labeled with a detectable marker is monoclonal antibody B503 or B504 produced by hybridomas B503 (ATCC No. 11999) and B504 (ATCC No. 12002) respectively. 
     
     
         31 . The method of  claim 30 , wherein the detectable marker is a radioactive isotope, enzyme, dye, magnetic bead, or biotin. 
     
     
         32 . The method of  claim 31 , wherein the detectable marker is the radioactive isotope is I 125 . 
     
     
         33 . A method for determining the efficacy of hormone replacement therapy in a perimenopausal female subject comprising the steps of:
 a. contacting a sample from the subject with an antibody which specifically binds to hLHβcf without substantially cross-reacting with hLH, hLHβ or hCGβcf, under conditions permitting formation of a complex between the antibody and hLHβcf;   b. measuring the amount of complex formed, so as to thereby determine the amount of hLHβcf, and   c. comparing the amount of hLHβcf measured in step (b) with either (i) the amount determined for a sample from a subject taken prior to the commencement of therapy or (ii) the amount determined for a sample after a prior course of therapy (iii) the amount determined for a sample from a known premenopausal female subject or (iv) the amount determined for a sample from a known postmenopausal female, wherein differences in the amounts of hLHβcf in the sample indicate efficacy of the hormone replacement therapy for the subject; amounts of hLHβcf in the sample similar to amounts of hLHβcf samples from known premenopausal subjects indicates efficacy of the hormone replacement therapy for the subject; amounts of hLHβcf molecule in the sample similar to amounts of hLβcf in the sample from known postmenopausal subjects indicates lack of efficacy of the hormone replacement therapy for the subject.   
     
     
         34 . The method of  claim 33 , wherein step (a) the capturing antibody specifically binds a region of hLHβcf comprising a protein portion or both a protein and a carbohydrate moiety. 
     
     
         35 . The method of  claim 33 , wherein in step (a) the antibody is bound to a solid support and in step (b) the amount of the antibody bound to the solid support in the complex with hlHβcf is measured by contacting the complex with a second antibody which binds to the complex and which is labeled with a detectable marker. 
     
     
         36 . The method of  claim 33 , wherein the sample is a urine sample, a first morning void urine sample, an aggregate sample of the first morning void urine samples for at least two consecutive days, or an aggregate sample of the first morning void urine samples for five or more consecutive days, or a collection of all urinations consecutively with time noted for 24-28 hours. 
     
     
         37 . The method of  claim 33 , wherein the replacement hormone therapy comprises therapy with estrogen. 
     
     
         38 . The method of  claim 33 , wherein the second antibody labeled with a detectable marker is monoclonal antibody B503 or B504 produced by hybridomas B503 (ATCC No. 11999) and B504 (ATCC No. 12002) respectively. 
     
     
         39 . The method of  claim 38 , wherein the detectable marker is a radioactive isotope, enzyme, dye, magnetic bead, or biotin. 
     
     
         40 . The method of  claim 39 , wherein the detectable marker is the radioactive isotope is I 125 . 
     
     
         41 - 45 . (canceled)