I. BF1H1 Cassette

• US 20030170646A1
• Filed: 04/19/2002
• Published: 09/11/2003
• Est. Priority Date: 04/20/2001
• Status: Active Grant

• Current Assignee: Azad Kaushik, Surinder Singh Saini
• Original Assignee: Azad Kaushik, Surinder Singh Saini
• Inventors: Saini, Surinder Singh, Kaushik, Azad
• Granted Patent: US 6,740,747 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]

Abstract

The present invention relates to a bovine VDJ cassette (BF1H1) that provides the novel ability to develop chimeric immunoglobulin molecule capable of incorporating both linear T cell epitope(s) (CDR1H and CDR2H) as well as conformational B cell epitope(s) (exceptionally long CDR3H). Further, multiple epitopes can be incorporated for development of multivalent vaccine by replacing at least a portion of an immunoglobulin molecule with the desired epitope such that functional ability of both epitope(s) and parent VDJ rearrangement is retained. The antigenized immunoglobulin incorporating both T and B epitopes of interest is especially useful for development of oral vaccines for use in humans apart from other species including cattle. The long CDR3H in BF1H1 VDJ rearrangement originates from long germline D-genes. The novel bovine germline D-genes provide additional opportunities for sustaining the capacity for antibody diversification in cattle essential for immunocompetence via selective breeding strategies that incorporate immunoglobulin gene markers. The novel gene elements, such as D-genes, are unique to cattle and, therefore, are useful in forensic analysis.

Claims

1. An isolated bovine VDJ gene cassette comprising the nucleic acid sequence shown in FIG. 1 (SEQ.ID.NO.1).

3. A use of a cassette according to claim 1 or the plasmid according to claim 2 to prepare a vaccine vector.

4. A use of a cassette according to claim 1 or the plasmid according to claim 2 in immunotherapy.

2. A recombinant plasmid pBF1H1-24 as shown in FIG. 1.

5. An isolated bovine germline D sequence comprising the nucleic acid sequence of D-1 (SEQ.ID.NO.2), D-2 (SEQ.ID.NO.3) and/or D-3 (SEQ.ID.NO.4) as shown in FIG. 2.

8. A use of a bovine D sequence according to claim 5 as a molecular marker.

11. A use of a bovine D sequence according to claim 5 as a DNA probe.

6. A recombinant plasmid pBGD14S as shown in FIG. 2.

9. A use of a plasmid according to claim 6 as a molecular marker.

12. A use of a plasmid according to claim 6 as a DNA probe.

7. A recombinant plasmid pBF1H1-D3 as shown in FIG. 3.

10. A use of a plasmid according to claim 7 as a molecular marker.

13. A use of a plasmid according to claim 7 as a DNA probe.