Method for the determination of gonadotropins
First Claim
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1. A method suitable for detection of luteinizing hormone in urine at a concentration as low as about 25 m.I.U./ml of urine and capable of detecting ovulation in a cycling woman, which comprises:
- (a) clarifying a sample of the urine to be tested;
(b) contacting the urine sample before, during or after clarification with 0.1 to 5% aqueous solution of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglobin, fibrinogen, human hemoglobin, and keyhold limpet hemomycin so that adsorption of luteinizing hormone from the sample onto the membrane is substantially reduced;
(c) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane having a molecular weight cut-off from about 5,000 to about 50,000, whereby luteinizing hormone is retained in the sample;
(d) continuing the ultrafiltration until the retentate sample is concentrated to one-tenth to one-five hundredth its original volume;
(e) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and
(f) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of luteinizing hormone in the retentate sample.
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Abstract
A simple, sensitive, reliable and safe method for determining the concentration of pituitary gonadotropins is disclosed. The method involves concentration by ultrafiltration of a sample of body fluid, e.g. urine or serum from a subject, followed by determining the presence of the gonadotropin in the concentrated sample.
20 Citations
12 Claims
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1. A method suitable for detection of luteinizing hormone in urine at a concentration as low as about 25 m.I.U./ml of urine and capable of detecting ovulation in a cycling woman, which comprises:
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(a) clarifying a sample of the urine to be tested; (b) contacting the urine sample before, during or after clarification with 0.1 to 5% aqueous solution of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglobin, fibrinogen, human hemoglobin, and keyhold limpet hemomycin so that adsorption of luteinizing hormone from the sample onto the membrane is substantially reduced; (c) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane having a molecular weight cut-off from about 5,000 to about 50,000, whereby luteinizing hormone is retained in the sample; (d) continuing the ultrafiltration until the retentate sample is concentrated to one-tenth to one-five hundredth its original volume; (e) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and (f) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of luteinizing hormone in the retentate sample. - View Dependent Claims (2)
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3. A method for detecting follicle stimulating hormone or human menopausal gonadotropin in urine at a concentration of 15 or more m.I.U./ml of urine, which comprises:
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(a) clarifying a sample of the urine to be tested; (b) contacting the urine sample before, during or after clarification with 0.1 to 5% aqueous solution of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglobin, fibrinogen, human hemoglobin, and keyhold limpet hemomycin so that adsorption of follicle-stimulating hormone or human menopausal gonadotropin from the sample onto the membrane is substantially reduced; (c) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane having a molecular weight cut-off from about 5,000 to about 50,000, whereby follicle-stimulating hormone or human menopausal gonadotropin is retained in the sample; (d) continuing the ultrafiltration until the retentate sample is one-tenth to one-five hundredth its original volume; (e) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and (f) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of follicle-stimulating hormone or human menopausal gonadotropin.
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4. A test method suitable for detection of pituitary gonadotropins in urine at a concentration as low as about 15 m.I.U./ml of urine which comprises:
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(a) clarifying a sample of the urine to be tested; (b) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane, said membrane having been prewashed with 0.1 to 5% aqueous solution of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglogin, fibrinogen, human hemoglobin, and keyhold limpet hemocyanin so that adsorption of the gonadotropin from the sample onto the membrane is substantially reduced, and said membrane having a molecular weight cut-off from about 5,000 to about 50,000 whereby the gonadotropin is retained in the sample; (c) continuing the ultrafiltration until the retentate sample is one-tenth to one-five hundredth its original volume; (d) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and (e) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of the gonadotropin in the retentate sample. - View Dependent Claims (5, 6, 7)
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8. A test method suitable for detection of pituitary gonadotropins in urine at a concentration as low as about 15 m.I.U./ml of urine which comprises:
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(a) clarifying a sample of the urine to be tested; (b) contacting the urine sample before or after clarification with a sufficient amount of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglobin, fibrinogen, human hemoglobin, and keyhold limpet hemocyanin to give a concentration of the protein in the urine of from 0.001 to 1.0% so that adsorption of the gonadotropin from the sample onto the membrane is substantially reduced; (c) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane having a molecular weight cut-off from about 5,000 to about 50,000 whereby the gonadotropin is retained in the sample; (d) continuing the ultrafiltration until the retentate sample is one-tenth to one-five hundredth its original volume; (e) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and (f) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of the gonadotropin in the retentate sample. - View Dependent Claims (9, 10)
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11. A test method suitable for detection of pituitary gonadotropins in urine at a concentration as low as about 15 m.I.U./ml of urine, which comprises:
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(a) clarifying a sample of the urine to be tested through a filter, said filter having been pretreated with an 0.1 to 5.0% aqueous solution of a protein selected from the group consisting of bovine serum albumin, rabbit serum albumin, human serum albumin, gamma globulin, myoglobin, fibrinogen, human hemoglobin, and keyhold limpet hemocyanin so that adsorption of the gonadotropin from the sample onto the membrane is substantially reduced; (b) subjecting about 5 to 50 ml of the clarified urine sample to absorbent induced ultrafiltration through an ultrafiltration membrane having a molecular weight cut-off from about 5,000 to about 50,000, whereby the gonadotropin is retained in the sample; (c) continuing the ultrafiltration until the retentate sample is one-tenth to one-five hundredth its original volume; (d) diluting the retentate sample with sufficient water to provide a retentate sample of about 0.5 ml; and (e) contacting the retentate sample with the appropriate amount of an immunological reagent for detecting the presence of the gonadotropin in the retentate sample. - View Dependent Claims (12)
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Specification