Mesangial cell-derived receptors for advanced glycosylation endproducts and uses thereof

• US 5,202,424 A
• Filed: 08/23/1991
• Issued: 04/13/1993
• Est. Priority Date: 03/19/1984
• Status: Expired due to Term

• Current Assignee: BBDX, Inc.
• Original Assignee: Rockefeller University
• Inventors: Cerami, Anthony, Vlassara, Helen
• Primary Examiner(s): Lacey, David L.
• Assistant Examiner(s): WANG, GIAN
• Application Number:
  US07/749,444
• Time in Patent Office: 599 Days
• Field of Search: 435/7.1, 435/7.9, 435/7.92, 435/7.93, 435/7.94, 536/18.7, 514/402, 514/635, 514/2, 424/9, 530/356, 530/362, 530/387, 530/395, 530/396, 530/397, 530/398, 530/399, 530/380
• US Class Current: 530/395
• CPC Class Codes: A61K 2039/6081 Albumin; Keyhole limpet haemocyanin [KLH]
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/155 Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
• CPC Class Codes: A61K 31/195 having an amino group
• CPC Class Codes: A61K 31/415 1,2-Diazoles
• CPC Class Codes: A61K 31/4178 not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
• CPC Class Codes: A61K 39/0005 Vertebrate antigens from snakes A61K39/38
• CPC Class Codes: A61K 8/64 Proteins; Peptides; Derivatives or degradation products thereof
• CPC Class Codes: A61Q 19/08 Anti-ageing preparations
• CPC Class Codes: C07D 405/14 containing three or more hetero rings
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: G01N 2333/922 Ribonucleases (RNAses); Deoxyribonucleases (DNAses)
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6842 : Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
• CPC Class Codes: G01N 33/6893 : related to diseases not provided for elsewhere

Abstract

The present invention relates to a method and associated agents for measuring the presence and amount of advanced glycosylation endproducts in cells and fluids. The methods take advantage of the existence of receptors and receptor complexes for AGEs and include receptor-containing ligands comprising whole mesangial and other cells, mesangial cellular fragments and protein extracts therefrom. Competitive assays, sandwich assays and assays involving AGE antisera are disclosed. Numerous diagnostic applications are defined and test kits are also contemplated.

Claims

1. An isolated receptor protein reactive with advanced glycosylation end products, characterized as follows:

(a) it has a molecular mass from 30 to 50 kD;

(b) it is reactive with AGE-BSA, AGE-RNAse, AGE-collagen I and AGE-BSA reduced with NaBH₄ and has a binding affinity of 2.0±

0.4×

10^-6 M^-1 (kD=500 nM);

(c) it is non-reactive with BSA, collagen I, RNAse or chemically synthesized FFI-BSA; and

(d) it is present on mesangial cell membranes prior to purification.

3. The protein of claim 1 or the receptor proteins of claim 2 labeled with a detectable label.

4. An isolated receptor protein reactive with advanced glycosylation end products in accordance with claim 1 having a molecular mass of about 50 kD.

5. An isolated receptor protein reactive with advanced glycosylation end products in accordance with claim 1 having a molecular mass of about 40 kD.

6. An isolated receptor protein reactive with advanced glycosylation end products in accordance with claim 1 having a molecular mass of about30-35 kD.

2. A plurality of isolated receptor proteins reactive with advanced glycosylation endproducts, characterized as follows:

(a) at least one of said proteins has a molecular mass from 30 to 50 kD;

(b) the plurality of proteins is derived from mammalian mesangial cells;

(c) the proteins are reactive with AGE-BSA, AGE-RNAse, AGE-collagen I and AGE-BSA reduced with NaBH₄, having a binding affinity of 2.0±

0.4×

10^-6 M^-1 (kD=500 nM);

(d) the proteins are non-reactive with BSA, collagen I, RNAse or chemically synthesized FFI-BSA; and

(e) the proteins are present on mesangial cell membranes prior to purification in an amount sufficient to bind about 3.0±

0.25×

10⁵ AGE-modified protein molecules per mesangial cell.