Nucleotide sequence encoding a 52 kDa Ro/SSA autoantigen

• US 6,111,088 A
• Filed: 09/16/1992
• Issued: 08/29/2000
• Est. Priority Date: 05/07/1990
• Status: Expired due to Fees

• Current Assignee: Oklahoma Medical Research Foundation
• Original Assignee: Oklahoma Medical Research Foundation
• Inventors: Frank, Mark Barton, Itoh, Kazuko
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): Siew, Jeffrey
• Application Number:
  US07/945,830
• Time in Patent Office: 2,904 Days
• Field of Search: 536/23.5, 536/24.1, 536/24.31, 435/6, 435/270, 435/252.3, 435/320.1, 530/358, 530/389.2
• US Class Current: 536/23.5
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/4713 Autoimmune diseases, e.g. Insulin-dependent diabetes mellitus, multiple sclerosis, rheumathoid arthritis, systemic lupus erythematosus; Autoantigens
• CPC Class Codes: C07K 16/18 against material from animals or humans

Abstract

Complementary DNA encoding a 52 kDa form of a protein present in the human Ro/SSA ribonucleoprotein complex has been cloned. A lambda gt11 cDNA library made from human thymocyte mRNA was screened with serum from a SLE patient and two immunoreactive clones were isolated. These clones reacted with other patient sera which had anti-52 kDa Ro/SSA antibodies and with affinity purified anti-52 kDa Ro/SSA antibodies. Moreover, affinity purified antibodies eluted from fusion proteins of the isolated clones reacted only with the 52 kDa protein of lymphocytes in the Western blot. Ro/SSA RNAs were also precipitated with these affinity purified antibodies, further confirming that the clones encode a 52 kDa Ro/SSA antigen. The sequence differs from the previously reported 60 kDa Ro/SSA gene.

Claims

1. An isolated nucleic acid molecule separate from the human genome encoding a Ro/SSA human autoantigen having a molecular weight by SDS-PAGE and Western blot of 52 kDa, or encoding an effective amount of the amino acid sequence of the Ro/SSA autoantigen to form an epitope immunoreactive with an anti-Ro/SSA autoantibody and which is isolated by a process comprising hybridizing under standard conditions to nucleic acid probes having the sequence set forth in FIG. 1 or portions thereof.

2. The molecule of claim 1 consisting essentially of a protein encoding fragment, wherein the protein encoding fragment is a fragment between nucleotide position number 39 and nucleotide position number 1466 of the sequence set forth in FIG. 1.

3. The molecule of claim 1 comprising the entire sequence set forth in FIG. 1.

4. The molecule of claim 1 labelled with a detectable label for use as a probe.

7. The nucleic acid molecule of claim 1 further comprising expression vector sequences for expression in prokaryotes or eukaryotes.

5. An isolated nucleic acid molecule separate from the human genome consisting of nucleotides 1 to 38 of the sequence set forth in FIG. 1.

6. An isolated nucleic acid molecule separate from the human genome consisting of nucleotides 1467 to 1843 of the sequence set forth in FIG. 1.