Method and composition for preserving antigens and nucleic acids and process for utilizing cytological material produced by same

• US 5,849,517 A
• Filed: 07/16/1996
• Issued: 12/15/1998
• Est. Priority Date: 05/08/1991
• Status: Expired due to Term

• Current Assignee: Streck Laboratories Inc.
• Original Assignee: Streck Laboratories Inc.
• Inventors: Ryan, Wayne L.
• Primary Examiner(s): MINNIFIELD, NITA M
• Application Number:
  US08/682,926
• Time in Patent Office: 882 Days
• Field of Search: 435/450.51, 435/40.5, 435/40.52, 436/16, 436/18, 436/17, 436/8
• US Class Current: 435/40.51
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 2001/307 non-toxic, no Hg, no formaldehyde
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/106664 Blood serum or blood plasma standard or control
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent

Abstract

A method and composition for fixing and stabilizing tissues, cells, and cell components such that the antigenic sites and nucleic acids are preserved is provided. The fixative employs a formaldehyde donor that is non-toxic, non-flammable, and that stabilizes the cell with minimal damage to and alteration of the cell morphology. The cell antigenic sites are left intact so that studies with monoclonal antibodies may be conducted. Vaccines and related immunotherapeutic methods utilizing antigens stabilized by the fixative of the present invention are also provided. Also disclosed is a method for developing a positive control for test reagents and for test instrumentation.

Claims

1. A method of preserving a tissue sample having one or more antigenic sites, comprising suspending said tissue sample in a solution comprising a fixative in an amount sufficient to preserve the antigenic sites, wherein said fixative is selected from the group consisting of imidazolidinyl urea, diazolidinyl urea, and mixtures thereof.

2. The method of claim 1, wherein the antigenic sites include cluster designation markers.

3. The method of claim 2, wherein said antigenic sites include CD4⁺ antigenic sites.

4. The method of claim 1, wherein the solution comprises from about 4 to about 6 percent by weight imidazolidinyl urea.

5. The method of claim 4, wherein the solution comprises about 5 percent by weight imidazolidinyl urea.

6. The method of claim 1, wherein the solution further comprises phosphate buffered saline.

7. The method of claim 1, wherein the solution further comprises an additive selected from the group consisting of ethylenediaminetetraacetic acid, polyethylene glycol, and mixtures thereof.

8. The method of claim 7, wherein the solution further comprises about 0.001 to about 0.2 percent by weight ethylenediaminetetraacetic acid.

9. The method of claim 7, wherein the solution comprises up to about 1 percent by weight polyethylene glycol.

10. The method of claim 1, wherein the solution comprises a mixture of imidazolidinyl urea and diazolidinyl urea.

11. The method of claim 10, wherein the solution comprises a total concentration of imidazolidinyl urea and diazolidinyl urea from about 4 percent to about 10 percent by weight wherein the weight ratio of imidazolidinyl urea to diazolidinyl urea is from about 10:

1 to about 1;

10.

12. The method of claim 1, wherein the tissue sample is selected from the group consisting of normal peripheral blood, bone marrow, lymph node, spleen, umbilical cord, and solid tissues.

13. The method of claim 1, wherein the tissue sample is an abnormal tissue sample selected from the group consisting of leukemias and solid tissue cancer.

14. A method of preserving a tissue sample having one or more nucleic acids comprising suspending said tissue sample in a solution comprising a fixative in an amount sufficient to preserve said nucleic acids, wherein said fixative is selected from the group consisting of imidazolidinyl urea, diazolidinyl urea, and mixtures thereof.

15. The method of claim 14, wherein the nucleic acid is RNA.

16. The method of claim 15, wherein the nucleic acid is viral RNA.

17. The method of claim 16, wherein the nucleic acid is HIV RNA.

18. The method of claim 14, wherein the nucleic acid is DNA.

19. The method of claim 14, wherein the solution comprises from about 1 to about 20 percent by weight diazolidinyl urea.

20. The method of claim 19, wherein the solution comprises about 10 percent by weight diazolidinyl urea.

21. The method of claim 14, wherein the solution further comprises phosphate buffered saline.

22. The method of claim 14, wherein the solution comprises a mixture of imidazolidinyl urea and diazolidinyl urea.

23. The method of claim 22, wherein the solution comprises a total concentration of diazolidinyl urea and imidazolidinyl urea from about 4 percent to about 10 percent by weight wherein the weight ratio of diazolidinyl urea to imidazolidinyl urea is from about 1:

1 to about 1;

10.

24. The method of claim 14, wherein the tissue sample is selected from the group consisting of normal peripheral blood, bone marrow, lymph node, spleen, umbilical cord, and solid tissues.

25. The method of claim 14, wherein the tissue sample is an abnormal tissue sample from the group consisting of leukemias and solid tissue cancers.