A rapid FPLC method for purification of the third component of human and guinea pig complement.

Abstract:

:A method is described for the purification of human and guinea pig C3 from small amounts of serum. This procedure requires only two steps--polyethylene glycol (PEG) precipitation and fast protein liquid chromatography (FPLC) Mono Q HR 10/10 ion exchange chromatography. The protocol takes less than two hours to complete and yields 4-6 mg of purified C3. Similar results, in terms of antigenic and functional recovery, were obtained for both human and guinea pig components. About 67% of C3 antigen was recovered from eluted fractions with fully preserved specific activity. Isolated C3 was over 95% pure as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography; this level of purity was confirmed by the absence of any observable contamination as assessed by immunoelectrophoresis using high titer anti-whole human serum. This method allows rapid and reproducible purification of fully active human or guinea pig C3 on a daily basis.

journal_name

J Immunol Methods

authors

Basta M,Hammer CH

doi

10.1016/0022-1759(91)90290-v

subject

Has Abstract

pub_date

1991-08-28 00:00:00

pages

39-44

issue

1

eissn

0022-1759

issn

1872-7905

pii

0022-1759(91)90290-V

journal_volume

142

pub_type

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