Korean J Immunol.  1999 Dec;21(4):303-309.

Production of CD44v6 Antibody Fragments and Comparision of Their Speciticities

Abstract

CD44v6 was known as tumor marker for tumor progression and metastasis in various kinds of carcinomas. The CD44v6 monoclonal antibody was produced by cell cultures or mouse ascite fluids using CD44v6 hybridoma cells, and its immunogloburin G (IgG) was purified by Protein A column. Using immobilized ficin and cysteine, the antibody fragment Fab was produced and purified by Protein A. Four CD44v6 scFv molecules were produced from the recombinant DNA and phage antibody technology and prurified by His-tag affinity chromatography. In order to inspect the function and specificity of each antibody molecule, western-blotting and ELISA against CD44v5-6 recombinant proteins and irnmunodetection in human ovarian carcinomas were estabilished. The results showed that immunodiagnosis did not distinguish the types of antibody fragments, but western-blotting and ELISA results did show some difference of their specificities and biological properties. These studies will contribute as a model study for the immunodiagnosis and therapy using the IgG, Fab and scFv of CD44v6 antibody to obtain the early detection of tumor progression and metastasis using immunoscintigraphy.

Keyword

CD44v6 antibody; IgG; Fab; scFv; Immunodiagnosis

MeSH Terms

Animals
Bacteriophages
Cell Culture Techniques
Chromatography, Affinity
Cysteine
DNA, Recombinant
Enzyme-Linked Immunosorbent Assay
Ficain
Humans
Hybridomas
Immunoglobulin Fragments*
Immunoglobulin G
Immunologic Tests
Mice
Neoplasm Metastasis
Recombinant Proteins
Sensitivity and Specificity
Staphylococcal Protein A
Cysteine
DNA, Recombinant
Ficain
Immunoglobulin Fragments
Immunoglobulin G
Recombinant Proteins
Staphylococcal Protein A
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