Monoclonal antibody to fibulin-1 generated by genetic immunization

J Cell Biochem. 2003 Jul 1;89(4):647-52. doi: 10.1002/jcb.10563.

Abstract

Fibulin-1 (Fbln-1) is an extracellular matrix (ECM) and plasma glycoprotein. Considering the growing evidence indicating that Fbln-1 plays a role in cancer we sought to develop monospecific antibodies to better facilitate further studies of the function of Fbln-1 in breast cancer. Using a plasmid expression vector encoding full-length human Fbln-1D as an immunogen and CpG oligodeoxyribonucleotides as adjuvant a monoclonal antibody (MAb) against Fbln-1 was produced. This MAb, designated MEM-2 was of IgM isotype and reacted with bacterially expressed Fbln-1. Furthermore, MEM-2 reacted with Fbln-1 expressed in the ECM released by cultured human breast carcinoma SKBR-3 cells in ELISA, and also with Fbln-1 present in SKBR-3 cell extract in immunoprecipitation and Western blotting. MEM-2 also reacted with Fbln-1 in human breast carcinoma specimens. These findings illustrate the utility of genetic immunization as a means of generating monoclonal antibodies to tumor-related ECM proteins. MEM-2 represents a useful new tool for the study of Fbln-1 in breast cancer.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adjuvants, Immunologic / genetics
  • Adjuvants, Immunologic / pharmacology
  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / isolation & purification
  • Breast Neoplasms / immunology
  • Breast Neoplasms / metabolism
  • Calcium-Binding Proteins / genetics*
  • Calcium-Binding Proteins / immunology*
  • Calcium-Binding Proteins / metabolism
  • Carcinoma, Squamous Cell / immunology
  • Carcinoma, Squamous Cell / ultrastructure
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / immunology
  • Extracellular Matrix Proteins / metabolism
  • Humans
  • Hybridomas / immunology
  • Immunoglobulin Isotypes / chemistry
  • Immunoglobulin Isotypes / immunology
  • Immunoglobulin M / chemistry
  • Immunoglobulin M / immunology
  • Immunologic Tests / methods
  • Mice
  • Oligodeoxyribonucleotides / genetics
  • Oligodeoxyribonucleotides / immunology
  • Oligodeoxyribonucleotides / pharmacology
  • Plasmids / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Tumor Cells, Cultured

Substances

  • Adjuvants, Immunologic
  • Antibodies, Monoclonal
  • Calcium-Binding Proteins
  • Extracellular Matrix Proteins
  • Immunoglobulin Isotypes
  • Immunoglobulin M
  • Oligodeoxyribonucleotides
  • Recombinant Proteins
  • fibulin