Production of CD44v6 Antibody Fragments and Comparision of Their Speciticities.
- Author:
Insook HAN
;
Seok Kil ZEON
;
Kwan Kyu PARK
- Publication Type:Original Article
- Keywords:
CD44v6 antibody;
IgG;
Fab;
scFv;
Immunodiagnosis
- MeSH:
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
- From:Korean Journal of Immunology
1999;21(4):303-309
- CountryRepublic of Korea
- Language:Korean
-
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.
