Preparative protein purification in a multi-compartment electrolyser with immobiline membranes
1989; Elsevier BV; Volume: 475; Issue: 2 Linguagem: Inglês
10.1016/s0021-9673(01)89684-9
ISSN1873-3778
AutoresPier Giorgio Righetti, Elisabeth Wenisch, Michel Faupel,
Tópico(s)Microbial Metabolic Engineering and Bioproduction
ResumoPreparative electrophoresis in gel phases, including isoelectric focusing, is characterized by low load limits (only a few milligrams of protein per millilitre of matrix), low recoveries (rarely exceeding 70%) and heavy contamination from neurotoxic gel material (unreacted monomers and ungrafted oligomers). A multi-compartment electrolyser is described based on the principle of highly buffering Immobiline membranes of well defined isoelectric points (pI). Each chamber is delimited by isoelectric membranes having pIs encompassing the pI value of the components of interest in the mixture to be separated. Such components are transported electrophoretically from chamber to chamber until they collect in the chamber defining their pI values. As the sample feed is kept in a number of reservoirs equal to the number of chambers in the electrolysers, at the end of the purification process each reservoir will contain, under ideal conditions, a homogeneous protein fraction. Examples of the purification of r-DNA Eglin C and of monoclonal antibodies are given.
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