On the domain pairing in chimeric antibodies
Alexey Teplyakov1, Galina Obmolova, Jill M Carton
1Centocor R&D, Inc., 145 King of Prussia Road, Radnor, PA 19087, USA. ateplyak@its.jnj.com
Molecular Immunology
|June 18, 2010
Summary
Researchers created a novel "double chimera" antibody by combining mouse variable regions with human and mouse constant regions. The study revealed stable antibody structure and preserved antigen-binding despite significant modifications, highlighting the plasticity of antibody constant domains.
Area of Science:
- Immunology
- Structural Biology
- Biochemistry
Background:
- Antibody engineering aims to create novel therapeutics with improved properties.
- Chimeric antibodies combine elements from different antibodies to achieve desired functions.
- Understanding the structural basis of antibody stability and function is crucial for rational design.
Purpose of the Study:
- To construct and characterize a novel "double chimera" antibody with mixed variable and constant regions.
- To determine the crystal structure of the chimeric antibody Fab fragment.
- To investigate the structural consequences of combining disparate antibody components on stability and antigen binding.
Main Methods:
- Construction of a chimeric antibody with mouse variable regions (targeting IL-13 and EMMPRIN) and mixed mouse/human constant regions.
- Expression of the chimeric antibody Fab fragment in mammalian cells.
- X-ray crystallography to determine the structure at 1.6Å resolution.
Main Results:
- The double chimera antibody formed a stable molecule despite numerous amino acid substitutions.
- The variable domain interface and antigen-binding loops remained conformationally preserved.
- Comparison with parent antibody structures revealed plasticity in the inter-chain interface of the constant domains.
Conclusions:
- The study demonstrates the feasibility of creating stable chimeric antibodies with mixed origins.
- Conservation of key residues maintains the integrity of the antigen-binding site.
- Antibody constant domains exhibit significant structural plasticity, allowing for tolerance of inter-chain interface modifications.
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