Related Experiment Videos
Expression of human recombinant CD23 in insect cells
K U Jansen1, J Shields, J Gordon
1Glaxo Institute for Molecular Biology S.A., Geneva, Switzerland.
Journal of Receptor Research
|January 1, 1991
Summary
Researchers successfully produced biologically active recombinant human CD23 (low affinity receptor for IgE) in insect cells using a baculovirus system. This provides a valuable source for studying CD23
Area of Science:
- Molecular Biology
- Immunology
- Biotechnology
Background:
- CD23, the low-affinity receptor for IgE, plays a crucial role in immune regulation.
- Understanding CD23's function requires a reliable source of recombinant protein.
Purpose of the Study:
- To express biologically active human CD23 in insect cells using the baculovirus system.
- To characterize the recombinant CD23 for its antigenic properties, IgE binding, and biological activity.
Main Methods:
- Baculovirus expression system utilized for human CD23 gene insertion into Sf9 insect cells.
- SDS-PAGE and Western blotting employed for molecular weight determination and antigenic analysis.
- IgE binding assays and B-cell growth promotion assays used to assess functional activity.
Main Results:
- Recombinant CD23 synthesized in insect cells exhibited antigenic properties similar to natural CD23.
- Surface expression confirmed by IgE binding; recombinant CD23 showed slightly lower molecular weight (43 kDa vs. 45 kDa).
- Affinity-purified recombinant CD23 demonstrated B-cell growth promoting activity.
Conclusions:
- The baculovirus expression system successfully produced biologically active recombinant human CD23.
- This recombinant CD23 is a valuable tool for further research into its immunological functions.
- Demonstrates the potential of insect cell expression systems for producing functional glycoproteins.