Purification of the First Component of Complement by Zonal Ultracentrifugation

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Abstract
The zonal centrifuge has been used to purify viruses, sub-cellular organelles and macromolecules of biologic interest (1, 2). In this report we describe a two-cycle preparative ultracentrifugation method for the separation from serum of chemically and functionally highly purified first component (C1) of guinea pig and human complement. The observation that the sedimentation rate of C1 varies as a function of ionic strength (3) made the development of this purification procedure possible. Materials and Methods. The source and preparation of sheep erythrocytes (E), rabbit antibody to boiled stromata of sheep erythrocytes (A) and sensitized sheep erythrocytes (EA) have been described (4). Fresh frozen guinea pig and human sera were purchased from Suburban Serum Laboratories, Silver Spring, Md. EAC4 were prepared with guinea pig serum as described (5). C1 activity was determined by the method described in (6).

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