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Purification of the third component of canine complement
Veterinary Immunology and Immunopathology
|April 1, 1985
Summary
A new method rapidly purifies the third component (C3) of canine complement from dog plasma. This technique yields highly pure C3, crucial for understanding complement system functions in dogs.
Area of Science:
- Immunology
- Biochemistry
Background:
- The complement system is vital for innate immunity.
- Canine complement component 3 (C3) plays a central role in complement activation.
- Efficient purification of canine C3 is essential for research.
Purpose of the Study:
- To develop a rapid and simple procedure for purifying canine complement component 3 (C3).
Main Methods:
- Fractionation of dog plasma using polyethylene glycol (PEG) precipitation.
- Plasminogen depletion using a Sepharose 4B-lysine column.
- Ion-exchange chromatography with DEAE-Sephacel and CM-Sepharose CL-6B columns.
Main Results:
- A single protein band of approximately 179 kDa was observed via SDS-PAGE.
- Reduced C3 showed two subunits (114 kDa and 65 kDa) by SDS-PAGE.
- Immunoelectrophoresis and antisera raised against purified C3 confirmed its purity.
Conclusions:
- The developed method provides a reliable way to obtain purified canine C3.
- This purified C3 can be used for further immunological and biochemical studies.
- The purification procedure is efficient, with a recovery of 18% for C3 antigen.
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