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Selected Publication:
Malle, E; Hess, H; Münscher, G; Knipping, G; Steinmetz, A.
Purification of serum amyloid A and its isoforms from human plasma by hydrophobic interaction chromatography and preparative isoelectric focusing.
Electrophoresis. 1992; 13(7):422-428
Doi: 10.1002/elps.1150130189
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- Leading authors Med Uni Graz
- Malle Ernst
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- Abstract:
- The present work was aimed at isolating human serum amyloid A, (SAA), an acute-phase protein mainly complexed to high density lipoproteins, directly from human plasma without sequential ultracentrifugation of lipoproteins and subsequent delipidation of the apolipoprotein moiety. Hydrophobic-interaction fast-protein liquid chromatography on Octylsepharose, using stepwise gradient elution profiles under dissociating conditions, followed by fast-protein liquid-gel permeation chromatography on a Superdex TM75 column revealed a higher than 95% purity of isolated SAA. Further purification of SAA from coeluting apolipoproteins C and A-II was achieved by preparative isoelectric focusing between pH5-7 using a Rotofor apparatus. Separation of the main SAA isoforms, SAA1 (pI 6.5) and SAA1 des-Arg (pI 6.0, lacking the N-terminal arginine), was achieved by anion-exchange fast-protein liquid chromatography on a Fractogel EMD DEAE 650-S column. The purity of the SAA1 and SAA1 des-Arg isoforms, thus isolated, was checked by immunochemical techniques and amino acid analysis. With the described method various SAA isoforms can be isolated, purified and separated directly from human plasma/serum without prior ultracentrifugation.
- Find related publications in this database (using NLM MeSH Indexing)
- Blotting, Western -
- Chromatography -
- Electrophoresis, Polyacrylamide Gel -
- Humans -
- Immunoelectrophoresis -
- Isoelectric Focusing -
- Serum Amyloid A Protein - isolation and purification