Resolution of a complex ionic mixture of an apparently homogenous protein preparation by preparative electrophoresis.
Preparations of recombinant envelope glycoprotein E2 of hepatitis C virus (r-HCV E2), found to be homogeneous by N-terminal amino acid sequencing and mass spectrometry, resolved into multiple ionic species (isoforms) when analysed by isoelectric focusing (IEF) gel electrophoresis in the p1 range of 3-10. These isoforms possessed pI values in the range of 4.5-8.2. The major isoform with p1 value of approximately 7.1 was separated from the rest of them by employing a method developed on Gradiflow BF 200, a device based on preparative electrophoresis. This isoform was adjudged to be homogenous by IEF and by native polyacrylamide gel electrophoresis (PAGE).
Biomolecular Research Institute, 343 Royal Parade, Parkville, Victoria 3052, Australia. ashokdubey@nsit.ac.in
Ashok K
AK
Dubey
D000180:Acrylic Resins / Q000737:chemistry; D000596:Amino Acids / Q000737:chemistry; D001671:Biochemistry / Q000379:methods; D045424:Complex Mixtures; D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D016174:Hepacivirus / Q000378:metabolism; D007477:Ions / Q000737:chemistry*; D007525:Isoelectric Focusing / Q000379:methods; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000378:metabolism; D000099166:Polyacrylamides
D000180:Acrylic Resins; D000596:Amino Acids; D045424:Complex Mixtures; D007477:Ions; D020033:Protein Isoforms; D011506:Proteins; D014759:Viral Envelope Proteins; D000099166:Polyacrylamides; C091015:glycoprotein E2, Hepatitis C virus