RESUMO
An adsorbent showing enhanced selectivity for the enzyme RNase A was prepared by a surface imprinting procedure based on metal coordination. A metal chelating monomer, N-(4-vinyl)-benzyl iminodiacetic acid, was polymerized onto methacrylate-derivatized silica particles in the presence of RNase A and metal ions. Lysozyme and RNase A were separated on the adsorbent used as stationary phase in high-performance liquid chromatography.
Assuntos
Muramidase/isolamento & purificação , Ribonuclease Pancreático/química , Ribonuclease Pancreático/isolamento & purificação , Adsorção , Animais , Bovinos , Quelantes/síntese química , Cromatografia Líquida de Alta Pressão , Iminoácidos/síntese química , Espectroscopia de Ressonância Magnética , Conformação Proteica , Estirenos/síntese química , Propriedades de SuperfícieRESUMO
New methods and applications in the separation of biomolecules are reviewed, with an emphasis on the large-scale recovery of proteins. Highlights include the advent of flow-through particles in perfusion chromatography, which allows for very high flow rates, while retaining a high chromatographic efficiency.
Assuntos
Proteínas Recombinantes/isolamento & purificação , Biotecnologia/métodos , Cromatografia , MembranasAssuntos
Carboidratos/isolamento & purificação , Cromatografia de Afinidade/métodos , Cromatografia Líquida de Alta Pressão/métodos , Glicoproteínas/isolamento & purificação , Nucleotídeos/isolamento & purificação , Proteínas/isolamento & purificação , Animais , Cromatografia de Afinidade/instrumentação , Cromatografia Líquida de Alta Pressão/instrumentação , Humanos , Indicadores e ReagentesRESUMO
A series of agarose gels, substituted with hydrophobic groups, has been synthesized and used for binding studies with the coloured model protein, phycoerythrin. The degrees of substitution for the derivatives can easily be estimated with proton magnetic resonance (NMR). It has been found that the capacity of the derivatives for phycoerythrin increases with increasing hydrophobicity of the substituent, degree of substitution and increasing ionic strength. For column experiments the degree of substitution should lie in the range 40-100 mmol substituent/mol galactose. When it is excessively high, the flow characteristics of the columns are unsatisfactory and difficulties to achieve complete desorption may arise.