A simplified method for purification of human C5a from citrated plasma

Area of Science:

• Biochemistry
• Immunology

Background:

• The fifth component of complement (C5) is cleaved to produce C5a, an 11,000 Da glycopeptide.
• C5a is a significant soluble mediator of inflammation.

Purpose of the Study:

• To develop a simplified immunoadsorption technique for purifying human C5a.
• To adapt the method for purifying radiolabeled 125I-C5a.

Main Methods:

• Isolation of human C5 fragments from zymosan-activated plasma via affinity chromatography.
• Concentration of C5 fragments using CM 52 cellulose.
• Purification to homogeneity by gel filtration on Sephadex G-75 in phosphate-buffered saline.

Main Results:

• The developed technique successfully purified human C5a to homogeneity.
• Purified human C5a exhibited characteristic immunochemical and biological activity.
• The method was adapted for the purification of 125I-C5a.

Conclusions:

• A simplified and effective method for purifying human C5a has been established.
• This technique provides a straightforward approach to obtaining biologically active C5a.
• The method is suitable for both native and radiolabeled C5a purification.