[Separation of recombinant proteins by chromatography on vesicular sorbents].
Vesicle chromatography, a recently developed method for separation of biomolecules, uses the vesicular packing (VP) material (clusters of microcapsules derived from plant cells), which was tested with respect to its application for the recombinant protein separation. Since VP has a well-defined separation limit, biomolecules are distributed in two separate peaks: large molecules are excluded and small molecules permeate through cell walls into the empty cell lumen. Recombinant proteins frequently form oligomers, which differ from monomers not only in size but also chemically and biologically. In the present study, separations of the recombinant proinsulin fusion protein oligomer and monomer, the recombinant human gamma-interferon monomer and dimer and recombinant tumour necrosis factor-alpha were investigated. For peak identification, the fractions and starting samples of the recombinant proteins were analysed by HPLC. The separations occurred without any sorption effects and with high efficiency and resolution of the protein peaks at a short column (10 cm). The VP is characterised by a high load ability, which favours the scale-up purification of the recombinant proteins. The combination of VP and HPLC is a considerable advance in biotechnology separation.
Kliushnichenko
V E
VE
Humboldt-University, Department of Biology, Berlin.
Vul'fson
A N
AN
Titel
C
C
Ehwald
R
R
Miroshnikov
A I
AI
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007371:Interferon-gamma / Q000302:isolation & purification*; D011384:Proinsulin / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000302:isolation & purification*
D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D011384:Proinsulin
Razdelenie rekombinantnykh belkov s pomoshch'iu khromatografii na vezikuliarnykh sorbentov.