Modified .beta..sub.2 -microglobulin

• US 5,175,113 A
• Filed: 03/29/1990
• Issued: 12/29/1992
• Est. Priority Date: 01/16/1986
• Status: Expired due to Fees

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Christensen, Mogens, Larsen, Flemming S., Nissen, Mogens H., Thim, Lars, Zeuthen, Jesper
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): Bidwell, Carol E.
• Application Number:
  US07/550,919
• Time in Patent Office: 1,006 Days
• Field of Search: 436/548, 436/547, 436/518, 436/529, 436/530, 436/804, 530/300, 530/324, 530/325, 530/326, 530/830, 530/834, 530/387, 530/388, 530/388.25, 530/387.9, 530/387.3, 530/388.85, 530/391.1, 530/391.3, 530/403, 930/260, 424/85.8
• US Class Current: 436/548
• 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 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2833 against MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay

Abstract

Substantially pure modified β₂ -microglobulin (mβ₂ m) of the formula I ##STR1## wherein R₁ is 24-amino acid residue, with the sequence Ile-Gln-Arg-Thr-Pro-Lys-Ile-Gln-Val-Tyr-Ser-Arg-His-Pro-Ala-Glu-Asn-Gly-Lys-Ser-Asn-Phe-Leu-Asn, R₂ is a 30-amino acid residue with the sequence Tyr-Val-Ser-Gly-Phe-His-Pro-Ser-Asp-Ile-Glu-Val-Asp-Leu-Leu-Lys-Asn-Gly-Glu-Arg-Ile-Gly-Lys-Val-Glu-His-Ser-Asp-Leu-Ser, R₃ is a 20-amino acid residue with the sequence Trp-Ser-Phe-Tyr-Leu-Leu-Tyr-Tyr-Glu-Phe-Thr-Pro-Thr-Glu-Lys-Asp-Glu-Tyr-Ala, R₄ is a 19-amino acid residue with the sequence Arg-Val-Asn-His-Val-Thr-Leu-Ser-Gln-Pro-Lys-Ile-Val-Lys-Trp-Asp-Arg-Asp-Met, X is Phe, Phe-Ser, or Phe-Ser-Lys, and Y is Asp, Lys-Asp, or Ser-Lys-Asp is disclosed. The presence of the protein in body fluids is a diagnostic and/or prognostic marker for the development of a variety of disorders such as different types of cancer and diseases involving the immune system. Also disclosed are specific anti-mβ₂ m antibodies used for the detection and quantification of mβ₂ m.

Claims

1. A monoclonal antibody that specifically binds to modified β

₂ -microglobulin of formula I wherein formula I is;

##STR4## wherein R₁ is a 24-amino acid residue with the sequence Ile-Gln-Arg-Thr-Pro-Lys-Ile-Gln-Val-Tyr-Ser-Arg-His-Pro-Ala-Glu-Asn-Gly-Lys-Ser-Asn-Phe-Leu-Asn, R₂ is a 30-amino acid residue with the sequence Tyr-Val-Ser-Gly-Phe-His-Pro-Ser-Asp-Ile-Glu-Val-Asp-Leu-Leu-Lys-Asn-Gly-Glu-Arg-Ile-Gly-Lys-Val-Glu-His-Ser-Asp-Leu-Ser, R₃ is a 20-amino acid residue with the sequence Trp-Ser-Phe-Tyr-Leu-Leu-Tyr-Tyr-Thr-Glu-Phe-Thr-Pro-Thr-Glu-Lys-Asp-Glu-Tyr-Ala, R₄ is a 19-amino acid residue with the sequence Arg-Val-Asn-His-Val-Thr-Leu-Ser-Gln-Pro-Lys-Ile-Val-Lys-Trp-Asp-Arg-Asp-Met, X is Phe, Phe-Ser, or Phe-Ser-Lys, and Y is Asp, Lys-Asp, or Ser-Lys-Asp, wherein said monoclonal antibody specifically binds to modified β

₂ -microglobulin but does not bind to β

₂ -microglobulin.

2. A monoclonal antibody according to claim 1 wherein the immunogen used for immunizing an animal is an antigen selected from the group consisting of mβ

₂ m, and compounds of the general formula II

space="preserve" listing-type="equation">R--Z II

wherein R is a carrier protein, and Z is a peptide of the formula R₂ '"'"'-X or R₃ '"'"'-Y wherein X is Phe, Phe-Ser, or Phe-Ser-Lys, and Y is Asp, Lys-Asp, or Ser-Lys-Asp, R₂ '"'"' is a peptide with from 1 to 30 amino acid residues corresponding to a fragment from the C-terminal end of R₂ wherein R2 is a 30-amino acid residue with the sequence Tyr-Val-Ser-Gly-Phe-His-Pro-Ser-Asp-Ile-Glu-Val-Asp-Leu-Leu-Lys-Asn-Gly-Glu-Arg-Ile-Gly-Lys-Val-Glu-His-Ser-Asp-Leu-Ser, and R₃ '"'"' is a peptide with from 1 to 20 amino acid residues corresponding to a fragment from the N-terminal end of R₃ wherein R₃ is a 20-amino acid residue with the sequence Trp-Ser-Phe-Tyr-Leu-Leu-Tyr-Tyr-Thr-Glu-Phe-Thr-Pro-Thr-Glu-Lys-Asp-Glu-Tyr-Ala.

3. The monoclonal antibody of claim 1 wherein said monoclonal antibody is a mouse monoclonal antibody.

4. The monoclonal antibody of claim 1 wherein said monoclonal antibody is detectably labeled.

5. The monoclonal antibody of claim 4 wherein said monoclonal antibody is radiolabeled.

6. The monoclonal antibody of claim 1 wherein said monoclonal antibody is bound to an insoluble solid phase.

7. A method for detecting modified β

₂ -microglobulin in human body fluid suspected of containing modified β

₂ -microglobulin, comprising contacting the body fluid with the monoclonal antibody of claim 1 detecting binding of said monoclonal antibody to mβ

₂ m in said body fluid, and evaluating the presence of said mβ

₂ m as a function of said binding of said monoclonal antibody to mβ

₂ m.

8. The method of claim 7 wherein said method is selected from the group consisting of dot-blot analysis, immunometric sandwich assay, and competitive assay.

9. A method for detecting modified β

₂ -microglobulin in human body fluid suspected of containing modified β

₂ microglobulin, comprising contacting the body fluid with the monoclonal antibody of claim 4 detecting binding of said monoclonal antibody to mβ

₂ m in said body fluid, and evaluating the presence of said mβ

₂ m as a function of said binding of said monoclonal antibody to mβ

₂ m.

10. The method of claim 9 wherein said method is selected from the group consisting of dot-blot analysis, immunometric sandwich assay, and competitive assay.

11. A method for detecting modified β

₂ -microglobulin in human body fluid suspected of containing modified β

₂ microglobulin, comprising contacting the body fluid with the monoclonal antibody of claim 6 detecting binding of said monoclonal antibody to mβ

₂ m in said body fluid, and evaluating the presence of said mβ

₂ m as a function of said binding of said monoclonal antibody to mβ

₂ m.

12. The method of claim 11 wherein said method is selected from the group consisting of dot-blot analysis, immunometric sandwich assay, and competitive assay.

13. The monoclonal antibody of claim 1, wherein X is Phe and Y is Asp.