Methods for determining antibodies specific for sex associated sperm membrane proteins

• US 5,660,997 A
• Filed: 06/07/1995
• Issued: 08/26/1997
• Est. Priority Date: 04/08/1987
• Status: Expired due to Fees

• Current Assignee: Infigen, Inc. (Astellas Pharma Incorporated)
• Original Assignee: Cytogam Incorporated
• Inventors: Spaulding, Glenn F.
• Primary Examiner(s): Hutzell, Paula K.
• Assistant Examiner(s): GRUN, JAMES LESLIE
• Application Number:
  US08/486,593
• Time in Patent Office: 811 Days
• Field of Search: 435/2, 435/7.21, 435/7.94, 435/7.95, 435/806, 436/503, 436/510, 436/547, 436/548, 436/518, 436/519, 436/65, 436/906, 530/350, 530/413, 530/388.85, 530/852, 600/35
• US Class Current: 435/7.21
• CPC Class Codes: A61K 35/52 Sperm; Prostate; Seminal fluid; Leydig cells of testes
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2800/367 Infertility, e.g. sperm disorder, ovulatory dysfunction
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads
• CPC Class Codes: Y10S 424/811 involving sex selection or contraception
• CPC Class Codes: Y10S 435/806 Fertility tests
• CPC Class Codes: Y10S 436/906 Fertility tests
• CPC Class Codes: Y10S 530/852 Sperm

Abstract

Substantially pure sex-associated membrane (SAM) proteins are used in methods to detect specific antibodies binding to X- or Y-SAM proteins.

Claims

1. A method of detecting antibodies to a sex-associated sperm membrane (SAM) protein in a sample comprising the steps of:

(a) contacting the sample with a substantially pure SAM protein, said SAM protein specifically binding to any antibodies to said SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex; and

(b) detecting said antibodies to said SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

2. A method of detecting antibodies to an X sex-associated sperm membrane (X-SAM) protein in a sample comprising the steps of:

(a) contacting the sample with a substantially pure X-SAM protein, said X-SAM protein specifically binding to any antibodies to said X-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex; and

(b) detecting said antibodies to said X-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

3. A method of detecting antibodies to a Y sex-associated sperm membrane (Y-SAM) protein in a sample comprising the steps of:

(a) contacting the sample with a substantially pure Y-SAM protein, said Y-SAM protein specifically binding to any antibodies to said Y-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex; and

(b) detecting said antibodies to said Y-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

4. A method of detecting antibodies to an X sex-associated membrane (X-SAM) protein in a simple comprising the steps of;

(a) contacting the sample with a substantially pure X-SAM protein, said X-SAM protein specifically binding to any antibodies to said X-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex, said X-SAM protein being characterized by a higher band density on an SDS/PAGE and a higher spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from X-enriched sperm subpopulations as compared to the corresponding band density on an SDS/PAGE and the corresponding spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from whole sperm or prepared from Y-enriched sperm subpopulations; and

(b) detecting said antibodies to said X-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

5. A method of detecting antibodies to a Y sex-associated membrane (Y-SAM) protein in a sample comprising the steps of;

(a) contacting the sample with a substantially pure Y-SAM protein, said Y-SAM protein specifically binding to any antibodies to said Y-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex, said Y-SAM protein being characterized by a higher band density on an SDS/PAGE and a higher spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from Y-enriched sperm subpopulations as compared to the corresponding band density on an SDS/PAGE and the corresponding spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from whole sperm or prepared from X-enriched sperm subpopulations; and

(b) detecting said antibodies to said Y-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

6. A method of detecting antibodies to an X sex-associated membrane (X-SAM) protein in a sample comprising the steps of;

(a) contacting the simple with a substantially pure X-SAM protein, said X-SAM protein specifically binding to any antibodies to said X-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex, said X-SAM protein being characterized by a higher band density on an SDS/PAGE and a higher spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from X-enriched sperm subpopulations as compared to the corresponding band density on an SDS/PAGE and the corresponding spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from whole sperm or prepared from Y-enriched sperm subpopulations and being selected from the group consisting of proteins having a molecular weight (KD) and pI as follows;

(1) 20.9, 5.74;

(2) 26.3, 7.58;

(3) 27.8, 6.08;

(4) 44.1, 6.90;

(5) 52.5, 5.33;

(6) 58.0, 5.99;

(7) 59.4, 6.59;

(8) 59.5, 6.81;

(9) 62.1, 7.23;

(10) 62.5, 5.54;

(11) 62.7, 6.85;

(12) 62.8, 6.64;

(13) 63.9, 5.83;

(14) 68.2, 5.95; and

(15) 78.6, 7.14; and

(b) detecting said antibodies to said X-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.

7. A method of detecting antibodies to a Y sex-associated membrane (Y-SAM) protein in a sample comprising the steps. of;

(a) contacting the sample with a substantially pure Y-SAM protein, said Y-SAM protein specifically binding to any antibodies to said Y-SAM protein in said sample to form a complex, under conditions and for a time sufficient to form said complex, said Y-SAM protein being characterizing by a higher band density on an SDS/PAGE and a higher spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from Y-enriched sperm subpopulations as compared to the corresponding band density on an SDS/PAGE and the corresponding spot density on an IPG-SDS/PAGE of plasma membrane proteins prepared from whole sperm or prepared from X-enriched sperm subpopulations and being selected from the group consisting of proteins having a molecular weight (KD) and pI as follows;

(1) 9.6, 6.58;

(2) 19.9, 5.67;

(3) 29.0, 6.67;

(4) 36.5, 7.16;

(5) 41.1, 6.21;

(6) 55.5, 6.82;

(7) 55.9, 5.25;

(8) 58.0, 8.67;

(9) 62.9, 6.34; and

(10) 30.3 5.77; and

(b) detecting said antibodies to said Y-SAM protein by contacting said complex with a detectable antibody which binds to immunoglobulin molecules.