Related Experiment Videos
Structural characterization of a recombinant CD4-IgG hybrid molecule
R J Harris1, K L Wagner, M W Spellman
1Department of Medicinal and Analytical Chemistry, Genentech, Inc., South San Francisco, CA 94080.
European Journal of Biochemistry
|December 12, 1990
Summary
This study characterizes CD4-IgG, an immunoadhesin combining CD4 binding with IgG properties. Structural analysis confirmed its expected features, including glycosylation and disulfide bonds.
Area of Science:
- Biochemistry
- Immunology
- Molecular Biology
Background:
- CD4 is a crucial co-receptor in T-cell activation.
- Immunoadhesins offer potential therapeutic advantages by combining binding specificity with effector functions.
Purpose of the Study:
- To structurally characterize CD4-IgG, an immunoadhesin.
- To confirm the fusion of human CD4 domains with human IgG-1 Fc domains.
- To validate the expected molecular features of this novel construct.
Main Methods:
- Peptide mapping using tryptic digestion.
- Analysis of tryptic peptides via chromatography.
- Deglycosylation and mass spectrometry to confirm glycosylation sites.
Main Results:
- Structural analysis confirmed 98.8% of the expected CD4-IgG structure.
- Presence of Asn-linked oligosaccharides at Asn257 was confirmed.
- Four intrachain and two interchain disulfide bonds were identified, consistent with IgG structure.
Conclusions:
- CD4-IgG possesses structural integrity mirroring both soluble CD4 and IgG Fc domains.
- The molecule's structure is well-defined, supporting its potential for therapeutic applications.
- The characterization provides a foundation for further functional studies of CD4-IgG.