Back to Search
Start Over
Isotype choice for chimeric antibodies affects binding properties.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 1994 Apr 29; Vol. 269 (17), pp. 13048-55. - Publication Year :
- 1994
-
Abstract
- Construction of a series of chimeric antibodies (murine variable region and human constant region) derived from the murine antibody BIRR1, which recognizes intercellular adhesion molecule 1 (ICAM-1), has revealed differences in the relative binding abilities of the chimeric antibody to antigen. The chimeric antibodies show a ranking of their ability to compete with BIRR1 for antigen on the surface of cells with the order BIRR1 = cIgG1 (100%) > cIgG4 (30%) > cIgG2 (10%) as demonstrated by solid-phase competitive enzyme-linked immunosorbent assay. Papain digestion yielded Fab fragments that were purified to homogeneity. Competitive enzyme-linked immunosorbent assay showed that the chimeric and murine Fab binding constants were equivalent. A solution-phase binding assay (analyzed by size exclusion high performance liquid chromatography) between the intact mAbs and recombinant soluble ICAM-1 further established that the binding constants involving the Fab arms of the two antibodies were equivalent. In summary, the murine and chimeric anti-ICAM-1 antibodies bind cellular ICAM-1 with equivalent affinities but with differing avidities.
- Subjects :
- Animals
Antibodies, Monoclonal genetics
Antibodies, Monoclonal isolation & purification
Base Sequence
CHO Cells
Cell Adhesion Molecules immunology
Cricetinae
Enzyme-Linked Immunosorbent Assay
Humans
Immunoglobulin Fab Fragments immunology
Immunoglobulin Isotypes genetics
Immunoglobulin Isotypes isolation & purification
Intercellular Adhesion Molecule-1
Mice
Molecular Sequence Data
Oligodeoxyribonucleotides
Recombinant Fusion Proteins
Antibodies, Monoclonal immunology
Binding Sites, Antibody
Immunoglobulin Isotypes immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0021-9258
- Volume :
- 269
- Issue :
- 17
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 7909805