Method of producing fibrin-specific monoclonal antibodies lacking fibrinogen-cross-reactivity using fibrin-specific peptides

• US 4,927,916 A
• Filed: 01/30/1986
• Issued: 05/22/1990
• Est. Priority Date: 04/23/1984
• Status: Expired due to Fees

• Current Assignee: General Hospital Corporation
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Haber, Edgar, Matsueda, Gary R., Hui, Kwan
• Primary Examiner(s): Moskowitz, Margaret
• Assistant Examiner(s): KUSHAN, JEFFREY
• Application Number:
  US06/824,228
• Time in Patent Office: 1,573 Days
• Field of Search: 530/387, 530/806, 530/382, 530/329, 435/240.26, 435/240.27, 435/172.2, 435/68, 935/93, 935/104, 935/95, 514/16
• US Class Current: 435/70.21
• CPC Class Codes: A61K 2123/00 Preparations for testing in vivo
• CPC Class Codes: A61K 51/1018 against material from animals or humans
• CPC Class Codes: A61K 51/1093 conjugates with carriers being antibodies
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins
• CPC Class Codes: Y10S 530/807 Hapten conjugated with peptide or protein
• CPC Class Codes: Y10S 530/808 Materials and products related to genetic engineering or hybrid or fused cell technology, e.g. hybridoma, monoclonal products
• CPC Class Codes: Y10S 530/809 Fused cells, e.g. hybridoma
• CPC Class Codes: Y10S 530/812 Peptides or proteins is immobilized on, or in, an organic carrier

Abstract

Peptides comprising fibrin-specific epitopic sequences are used to prepare hybridoma cell lines producing antifibrin-specific monoclonal antibodies substantially devoid of fibrinogen-cross-reactivity obtained by somatic cell fusion. The antibodies are useful for the in vivo and in vitro detection of thrombi and fibrin deposits.

Claims

1. A process for producing monoclonal anti-fibrin specific antibodies, devoid of cross reactivity with fibrinogen comprising:

a. obtaining lymphocytes from a animal immunized against the peptide fragment comprising

space="preserve" listing-type="equation">A-B-C-D-E-F-G

attached to a suitable immunogenic carrier, wherein

A is glycine;

B is histidine or proline;

C is arginine;

D is proline or valine;

E is aspartic acid or glutamic acid;

G is lysine or arginine;

b. fusing said lymphocytes with a meyloma cell line to produce hydridomas which secrete anti-fibrin specific antibodies substantially devoid of fibrinogen cross-reactivity;

c. cultivating said hybridomas; and

d. isolating said secreted antibodies.

2. The process of claim 1, wherein said lymphocytes are comprised of spleen cells.

3. The process of claim 2, wherein said spleen cells are mice spleen cells.