Expression of porcine reproductive respiratory syndrome virus polypeptides in the same cell

• US 5,858,729 A
• Filed: 03/14/1996
• Issued: 01/12/1999
• Est. Priority Date: 03/14/1995
• Status: Expired due to Term

• Current Assignee: Intervet International BV (Merck & Co., Inc.)
• Original Assignee: Akzo Nobel N.V. (Nouryon BV)
• Inventors: Conzelmann, Karl-Klaus, Van Woensel, Petrus Alphonsus Maria
• Primary Examiner(s): Knode, Marian C.
• Assistant Examiner(s): LEE, DATQUAN HUU
• Application Number:
  US08/616,032
• Time in Patent Office: 1,034 Days
• Field of Search: 435/320.1, 435/236, 435/237, 435/239, 435/235.1, 435/5, 435/173.3, 435/69.1, 435/71.1, 435/69.3, 530/395, 530/403, 935/32, 424/185.1, 424/204.1, 424/199.1
• US Class Current: 435/71.1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2710/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2770/10022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The present invention provides a process for the recombinant DNA production of a mature form of the PRRSV ORF 3 or ORF 4 protein. This is achieved by the co-expression of the PRRSV ORF 2, ORF 3 and ORF 4 proteins by the same cell.

Claims

1. A process for the preparation of a glycosylated forms of porcine reproductive respiratory syndrome virus (PRRSV) ORF 3 and ORF 4 proteins, comprising the steps of:

a. culturing a suitable host cell transformed with subgenomic DNA fragments encoding the PRRSV ORF 2, ORF 3 and ORF 4 proteins, or a suitable host cell infected with one or more vector (virus) viruses each of which comprise one or more subgenomic DNA fragments encoding the PRRSV ORF 2, ORF 3 or ORF 4 proteins, under conditions whereby the ORF 2, ORF 3 and ORF 4 proteins are expressed; and

b. harvesting the expressed proteins.

2. The process according to claim 1, wherein said host cell comprises PRRSV ORF 2, ORF 3 and ORF 4 as a result of coinfection with more than one vector virus.

3. The process according to claim 2, wherein the vector virus is a baculovirus.

10. An immunogenic composition comprising the ORF proteins produced by the process according to claim 1 together with a pharmaceutically acceptable carrier or diluent.

4. A host cell transformed with subgenomic DNA fragments encoding PRRSV ORF 2, ORF 3 and ORF 4.

5. A vector virus comprising two or more heterologous DNA fragments selected from the group consisting of the DNA fragments that encode PRRSV ORF 2, ORF 3 and ORF 4 proteins.

6. The vector virus according to claim 5, which is pseudorabies virus.

7. A mixture of two or more vector viruses, each vector virus comprising one or more subgenomic PRRSV ORFs, said mixture being able to co-infect a host cell and to express PRRSV ORF 2, ORF 3 and ORF 4 in the cell.

8. An isolated DNA molecule comprising a subgenomic nucleic acid sequence encoding PRRSV ORF 2, ORF 3 and ORF 4 proteins, each ORF being separately and operatively linked to a promoter.

9. An isolated, glycosylated, endo-H resistant form of PRRSV ORF 3 or ORF 4 protein.

11. An immunogenic composition comprising at least one endo-H resistant PRRSV protein selected from the group consisting of ORF 3 and ORF 4, together with a pharmaceutically acceptable carrier or diluent.

12. An immunogenic composition comprising at least one vector virus that singly or together will express ORF 2, ORF 3 and ORF 4 proteins in a cell, together with a pharmaceutically acceptable carrier or diluent.