Rapid isolation of human complement component C9 to verify the specificity of a haemolytic C9 microassay.
A sensitive, haemolytic microassay of human complement component C9 was developed. The assay is based on the principle of reactive (C5b6-initiated) haemolysis and uses commercially available C9-depleted serum as reagent for C9. The specificity of the assay was verified by rapid, activity-guided isolation of the haemolytic component from human serum using high-performance liquid chromatography (HPLC) on a system for fast protein liquid chromatography. This isolation yielded a single component with characteristics of C9. The results suggest that rapid, activity-guided isolation as a new application of HPLC can be a useful tool to demonstrate the specificity of a functional assay.
Bootsma
H J
HJ
Eijkman-Winkler Laboratory of Medical Microbiology, University Hospital, Utrecht, Netherlands.
van den Berg
C W
CW
van Dijk
H
H
D000328:Adult; D000818:Animals; D002645:Chickens; D002853:Chromatography, Liquid; D003186:Complement C9 / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D012680:Sensitivity and Specificity
D003186:Complement C9