MN Gene and Protein
A new gene—MN—and proteins/polypeptides encoded therefrom are disclosed. Recombinant nucleic acid molecules for expressing MN proteins/polypeptides and recombinant proteins are provided. Expression of the MN gene is disclosed as being associated with tumorigenicity, and the invention concerns methods and compositions for detecting and/or quantitating MN antigen and/or MN-specific antibodies in vertebrate samples that are diagnostic/prognostic for neoplastic and pre-neoplastic disease. Test kits embodying the immunoassays of this invention are provided. MN-specific antibodies are disclosed that can be used diagnostically/prognostically, therapeutically, for imaging, and/or for affinity purification of MN proteins/polypeptides. Also provided are nucleic acid probes for the MN gene as well as test kits comprising said probes. The invention also concerns vaccines comprising MN proteins/polypeptides which are effective to immunize a vertebrate against neoplastic diseases associated with the expression of MN proteins. The invention still further concerns antisense nucleic acid sequences that can be used to inhibit MN gene expression.
1 . An isolated nucleic acid containing at least fifty nucleotides wherein the nucleotide sequence for said nucleic acid is selected from the group consisting of:
(a) SEQ. ID. NOS.: 1, 5, 23 and nucleotide sequences complementary to SEQ. ID. NOS.: 1, 5 or 23;
(b) nucleotide sequences that hybridize under standard stringent hybridization conditions to one or more of the following nucleotide sequences: SEQ. ID. NOS.: 1, 5, 23 and the respective complements of SEQ. ID. NOS.: 1, 5 and 23; and
(c) nucleotide sequences that but for the degeneracy of the genetic code would hybridize under standard stringent hybridization conditions to one or more of the following nucleotide sequences: SEQ. ID. NOS.: 1, 5, 23 and the respective complements of SEQ. ID. NOS.: 1, 5 and 23.
2 . An isolated nucleic acid according to claim 1 which encodes an MN protein or polypeptide.
3 . An isolated nucleic acid according to claim 1 wherein the nucleotide sequence for said nucleic acid is selected from the group consisting of:
(a) SEQ. ID. NO.: 23 and its complement;
(b) nucleotide sequences that hybridize under standard stringent hybridization conditions to SEQ. ID. NO.: 23 or to its complement; and
(c) nucleotide sequences that but for the degeneracy of the genetic code would hybridize under standard stringent hybridization conditions to SEQ. ID. NO.: 23 or to its complement;
wherein the nucleic acids represented by the nucleotide sequences of (b) and (c) function to identify MN nucleic acid sequences.
4 - 5 . (canceled)
6 . An isolated nucleic acid according to claim 1 operatively linked to an expression control sequence within a vector.
7 . A unicellular host which is either prokaryotic or eukaryotic that is transformed or transfected with the isolated nucleic acid operatively linked to an expression control sequence in a vector according to claim 6 .
8 . A unicellular host according to claim 7 which is an insect cell or an E. coli cell.
9 . An MN protein or polypeptide encoded by the isolated nucleic acid according to claim 2 .
10 . A fusion protein comprising at least a first and a second amino acid wherein the first amino acid is encoded by an isolated nucleic acid according to claim 2 , and wherein the second amino acid is a non-MN protein or polypeptide.
11 . (canceled)
12 . A method of recombinantly producing an MN protein or polypeptide wherein a baculovirus expression system is used comprising the steps of:
(a) transforming an appropriate unicellular host with an isolated nucleic acid operatively linked to an expression control sequence in a vector according to claim 6 ;
(b) culturing said unicellular host so that said MN protein or polypeptide is expressed; and
(c) extracting and isolating said MN protein or polypeptide.
13 . An antibody which specifically binds to an epitope of an MN protein or polypeptide according to claim 9 .
14 . An antibody according to claim 13 which is monoclonal.
15 - 16 . (canceled)
17 . An antibody according to claim 13 which specifically binds to an MN antigen epitope selected from the group of epitopes represented by the following amino acid sequences: SEQ. ID. NOS. 10-16.
18 - 21 . (canceled)
22 . An antibody according to claim 13 which is linked to a chemotherapeutic agent or a toxic agent.
23 . A method of delivering a chemotherapeutic agent or toxic agent to a cancer cell which comprises contacting said cell with an antibody according to claim 22 .
24 . An antibody according to claim 13 which is linked to an imaging agent.
25 . A method of imaging pre-neoplastic or neoplastic disease in a patient comprising:
(a) injecting said patient with antibody according to claim 24 ; and
(b) detecting the binding of said antibody.
26 . A method of detecting and/or quantitating in a vertebrate sample MN antigen comprising the steps of:
(a) contacting said sample with an antibody according to claim 13 ; and
(b) detecting and/or quantitating binding of said antibody in said sample.
27 . The method according to claim 26 wherein said detecting and/or quantitating is by immunohistochemical staining.
28 . The method according to claim 26 which is diagnostic/prognostic for pre-neoplastic/neoplastic disease.
29 . A method of detecting and/or quantitating MN-specific antibodies in a vertebrate sample comprising the steps of:
(a) contacting and incubating the vertebrate sample with a fusion protein according to claim 10 ; and
(b) detecting and/or quantitating binding of said fusion protein to antibody in said sample.
30 . (canceled)