Idiotype and anti-idiotype antibodies useful in virus detection

• US 4,910,131 A
• Filed: 12/23/1987
• Issued: 03/20/1990
• Est. Priority Date: 12/23/1987
• Status: Expired due to Fees

• Current Assignee: Yale University
• Original Assignee: Ari H. Helenius, David Vaux, Ira S. Mellman
• Inventors: Vaux, David, Mellman, Ira S., Helenius, Ari H.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US07/137,237
• Time in Patent Office: 818 Days
• Field of Search: 435/5, 435/7, 435/192.2, 435/235, 435/948, 436/548, 436/63, 530/387, 530/808, 530/809, 530/826
• US Class Current: 435/5
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: G01N 33/686 Anti-idiotype
• CPC Class Codes: Y10S 435/948 using viruses or cell lines
• CPC Class Codes: Y10S 530/808 Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
• CPC Class Codes: Y10S 530/809 Fused cells, e.g. hybridoma
• CPC Class Codes: Y10S 530/826 Viruses

Abstract

The invention describes a new process for the production of anti-viral diagnostics, as well as an example of one antibody useful in detecting a wide variety of pathogenically important alphaviruses. The process includes, but is not limited to, the isolation of monoclonal antibodies directed against internal viral components. Such antibodies may be generated by conventional immunization in vivo or in vitro using subviral particles or synthetic peptides corresponding to specific viral proteins. Alternatively, synthetic peptides derived from the cytoplasmic domains of envelope proteins may be used to elicit monoclonal idiotypic and anti-idiotypic antibodies following immunization in vitro and/or in vivo.

The anti-alphavirus antibody included in the invention is an example of the type of reagent produced. It was produced as an anti-idiotype against monoclonal antibodies to a synthetic peptide derived from Semliki Forest virus. The monoclonal anti-idiotype is specific for the genome-containing nucleocapsid structure not only of Semliki Forest virus, but of every alphavirus tested thus far.

Claims

1. A method for producing an antibody directed against highly conserved domains of viral components involved in virus assembly or stabilization of internal virus structure comprising the steps of:

(a) generating a polyclonal response by in vitro immunization to an antigenic component corresponding to a viral component involved in virus assembly or stabilization of internal virus structure, said response comprising polyclonal antibodies;

(b) using said polyclonal antibodies as an antigen to produce an internal image anti-idiotype antibody by in vitro immunization, said internal image antibody recognizing and binding to a highly conserved domain of a virus which acts as binding site for said antigenic component.

2. The method of claim 1, wherein said highly conserved domain is a functional domain.

3. The method of claim 1, wherein said internal image anti-idiotype antibody recognizes a highly conserved domain of an alphavirus.

4. The method of claim 3, wherein the internal image anti-idiotype antibody is specific for the nucleocapsid of said alphaviruses.

5. The method of claim 1, wherein said polyclonal response is generated in response to a synthetic peptide corresponding to a highly conserved domain of a viral component involved in virus assembly or stabilization of internal virus structure.

6. The method of claim 5, wherein said polyclonal response is generated in response to the following synthetic peptide corresponding to the SFV E2 cytoplasmic domain:

NH₂ -Arg-Ser-Lys-Cys-Leu-Thr-Pro-Try-Ala-Leu-Thr-Pro-Gly-Ala-Ala-Val-Pro-Trp-Try-Leu-Gly-Iso-Leu-Cys-Cys-Ala-Pro-Arg-Ala-His-Ala-COOH.

7. A method for detecting the presence of an enveloped virus comprising:

(a) generating a polyclonal response by in vitro immunization to an antigenic component corresponding to a conserved cytoplasmic domain of a virus spike glycoprotein, said response comprising polyclonal antibodies;

(b) using said polyclonal antibodies as an antigen to produce an internal image anti-idiotype antibody by in vitro immunization, said internal image anti-idiotype antibody recognizing and binding to a highly conserved functional domain of a virus which acts as a binding site for said antigenic component; and

(c) detecting said virus in an immunoassay.

8. The method of claim 7, wherein said immunoassay is an indirect immunofluorescence assay.

9. The method of claim 7, wherein said immunoassay is an ELISA assay.

10. The method of claim 7, wherein said immunoassay is an immunoprecipitation assay.

11. A method for detecting presence of a virus comprising:

(a) generating a polyclonal response by in vitro immunization to an antigenic component corresponding to a highly conserved domain of a viral component involved in virus assembly or stabilization of internal virus structure, said response comprising polyclonal antibodies;

(b) using said polyclonal antibodies as an antigen to produce an internal image anti-idiotype antibody by in vitro immunization, said internal image anti-idiotype antibody recognizing and binding to a highly conserved domain of a virus which acts as a binding site for said antigenic component; and

(c) detecting said virus in an immunoassay.

12. A method for detecting the presence of an alphavirus comprising:

(a) generating a polyclonal response to a synthetic peptide corresponding to the highly conserved cytoplasmic domain of E2 glycoprotein of Semliki Forest virus in vitro, said response comprising polyclonal antibodies;

(b) using said polyclonal antibodies as an antigen to produce an internal image anti-idiotype monoclonal antibody in vitro, said internal image anti-idiotype monoclonal antibody recognizing and binding to a highly conserved functional domain of an alphavirus which acts as a binding site for the cytoplasmic domain of E2 glycoprotein; and

(c) detecting said alphavirus in an immunoassay.

13. The method of claim 12, wherein said internal image anti-idiotype antibody is generated in response to polyclonal antibodies provided in vitro against the following synthetic peptide corresponding to the SFV E2 cytoplasmic domain:

NH₂ -Arg-Ser-Lys-Cys-Leu-Thr-Pro-Try-Ala-Leu-Thr-Pro-Gly-Ala-Ala-Val-Pro-Trp-Try-Leu-Gly-Iso-Leu-Cys-Cys-Ala-Pro-Arg-Ala-His-Ala-COOH.

14. The method of claim 12, wherein said immunoassay is an immunofluorescence assay.

15. The method of claim 12, wherein said assay is an immunoprecipitation assay.

16. The method of claim 12, wherein said assay is an ELISA assay.

17. The method of claim 12, wherein said internal image anti-idiotype monoclonal antibody is IgM F13.3C5, produced by continuous cell line of ATCC Deposit No. HB9617.

18. An internal image anti-idiotype monoclonal antibody which binds to an epitope corresponding to a highly conserved functional domain of a virus nucleocapsid structure.

19. The monoclonal antibody of claim 18, wherein said virus is an alphavirus.

20. The monoclonal antibody of claim 19, wherein said antibody is anti-idiotype internal image monoclonal IgM F13.3C5 produced by continuous cell line of ATCC Deposit No. HB9617.

21. The continuous cell line of ATCC Deposit No. HB9617.