Isolation of late complement components by affinity chromatography. II. Purification of the human complement...

Area of Science:

• Immunology
• Biochemistry

Background:

• The human complement system is vital for innate immunity.
• Efficient isolation of complement components is essential for research.
• Previous methods for complement component isolation were limited.

Purpose of the Study:

• To develop a rapid and gentle procedure for human complement component C6 isolation.
• To achieve high purity and recovery of functional C6.

Main Methods:

• Affinity chromatography using insolubilized anti-C6 antibodies.
• Elution of C6 with 3 M thiocyanate, pH 7.2.
• Secondary purification using an "anti-impurity" column.

Main Results:

• Achieved 15-23% recovery and >270-fold purification of C6 in the first step.
• Final product showed 12% recovery and >1300-fold purification.
• The isolated C6 was homogeneous by SDS-PAGE and immunoelectrophoresis.

Conclusions:

• The developed affinity chromatography procedure is effective for rapid and gentle human C6 isolation.
• This method provides a highly purified and functional complement component C6.
• The technique is comparable to previous methods for C9 isolation.