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C1q: Rapid Purification Method for Preparation of Monospecific Antisera and for Biochemical Studies^1,2,

From the Division of Clinical Immunology and Rheumatology, Department of Medicine, The Medical Center, University of Alabama in Birmingham, 1919 Seventh Avenue, South Birmingham, Alabama 35233

Abstract

C1q has been purified using precipitation in the presence of chelating agents at low ionic strength. The three times precipitated C1q was highly purified as shown by immunoelectrophoresis and analytic acrylamide disc electrophoresis. On immunoelectrophoresis, the purified C1q showed a single line in the slow -region against a potent anti-whole human antiserum, and the hemolytic activity and latex agglutination activity of C1q coincided with this region.

The final product showed hemolytic activity and latex agglutination activity but did not show any detectable C1r or C1s activity. The yield of C1q using these methods is about 40 to 60% and thus about 5 mg can be obtained from 100 ml of serum. The highly purified C1q obtained from 3.75% preparative acrylamide gels containing sodium dodecyl sulfate yields potent monospecific antisera when injected into rabbits. These antisera agglutinate EAC1q cells but not EA. The molecular weight of C1q as estimated by its relative mobility on acrylamide gels was 387,600 ± 10,790, and breakdown to smaller subunits has been demonstrated in gels containing urea.

Footnotes

¹ This work was supported by grants from the National Institutes of Health, United States Public Health Service, and from the Veterans Administration.

² The symbols for complement components used in this paper conform to the recommendations of a committee sponsored by the World Health Organization. "Nomenclature of Complement," Immunochemistry, 7: 137, 1970. Complement (C) components are designated numerically C1, C2, C3, C4, C5, C6, C7, C8 and C9; the subunits of C1 are designated C1q, C1r and C1s; activated components are indicated by placing a bar over the numeral which refers to the component or subunit, i.e., active C1 = C; active C1s = Cs. As guinea pig components are always used for the C1 subunits which were from human serum, a superscript indicating the species will be omitted. Cellular intermediates carrying C components are designated EAC, followed by the numeral designating the components carried, e.g., EAC4.