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Depletion of Specific Cell Populations by Complement Depletion
Published on: February 5, 2010
Depletion of specific cell populations by complement depletion.
1BloodCenter of Wisconsin, Blood Research Institute, USA. bonnie.dittel@bcw.edu
Journal of Visualized Experiments : Jove
|February 9, 2010
Summary
Complement depletion is a fast, cost-effective method for purifying immune cells. This technique uses antigen-specific antibodies and rabbit complement to achieve high purity for molecular studies.
Area of Science:
- Immunology
- Cell Biology
Background:
- Immune cell purification is crucial for studying cellular functions.
- Molecular techniques like PCR and microarrays demand highly pure cell populations.
Purpose of the Study:
- To highlight complement depletion as an efficient cell purification strategy.
- To compare complement depletion with other methods like FACS and magnetic bead separation.
Main Methods:
- Utilizing complement-fixing antigen-specific monoclonal antibodies (mAb) and rabbit complement.
- Incubating cells with reagents for one hour at 37°C.
- Removing lysed cells through two washing steps.
Main Results:
- Complement depletion is fast, inexpensive, gentle, and yields high cell numbers.
- Monoclonal antibodies with efficient complement fixation can deplete 95-100% of targeted cells.
- This method can be used for direct purification or enrichment prior to further purification.
Conclusions:
- Complement depletion is a valuable technique for immune cell purification.
- It offers significant advantages over other methods for specific research applications.
- This method supports both standalone purification and enrichment strategies.
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