Isolation of capsid proteins of foot-and-mouth disease virus by chromatofocusing.
A method for the isolation of foot-and-mouth disease virus (FMDV) capsid proteins was developed. The FMDV capsid proteins VP1, VP2, VP3 and VP0 were isolated from sucrose gradient purified virus by chromatofocusing in a pH 7.4-4.0 gradient on Polybuffer exchanger PBE 94. Under the conditions used the proteins eluted in the sequence VP1, VP2, VP0 (when present) and VP3. Capsid protein VP4 did not elute and could not be isolated by this method. Protein concentration in the eluate was monitored by the use of a radiolabelled marker and recoveries of approximately 50% of the input marker could be achieved when using up to 15 mg of virus and a 30-ml column. The high capacity and relative simplicity of chromatofocusing make it a useful alternative to other methods of purifying proteins.
D000818:Animals; D005537:Aphthovirus / Q000032:analysis*; D002213:Capsid / Q000302:isolation & purification*; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing / Q000379:methods