C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
C07K 1/14 (2006.01)
Patent
CA 2699529
The object of the present invention is a procedure for the distribution, separation and purification in aqueous solution of recombinant proteins, based on the utilization of polypeptides with choline affinity. The invention is based on a phenomenon consisting of that two aqueous solutions with determinated components can be mixed, being distributed finally in two phases with different density. The fusionated proteins to said polypeptides with choline affinity are preferably located in one of the phases, while most of the cell extract proteins tend to go to the opposite phase. After a serie of washings for removing the rest of the not desired material, this location can be inverted through the addition of a soluble molecule with affinity by the polypeptide fusionated to the protein of interest. This. procedure allows modulating at convenience the presence of the protein or polypeptide of interest in one phase or another, possibiliting its purification with a high yield and purity grade. The invention represents an economic and scalable way of recombinant protein separation labelled preferably with choline-binding domains.
El objeto de la presente invención es un procedimiento para el reparto, separación y purificación en solución acuosa de proteínas recombinantes, basado en la utilización de polipéptidos con afinidad por colina. La invención se fundamenta en un fenómeno consistente en que dos soluciones acuosas con determinados componentes pueden ser mezcladas repartiéndose finalmente en dos fases con distinta densidad. Las proteínas fusionadas a dichos polipéptidos con afinidad por colina se localizan preferentemente en una de las fases, mientras que la mayoría de las proteínas de un extracto celular tienden a irse a la fase contraria. Tras una serie de lavados para eliminar el resto de material no deseado, esta localización puede invertirse mediante la adición de una molécula soluble con afinidad por el polipéptido fusionado a la proteína de interés. Este procedimiento permite modular a conveniencia la presencia de la proteína o polipéptido de interés en una fase u otra, posibilitando su purificación con un alto rendimiento y grado de pureza. La invención representa una forma económica y escalable de separación de proteínas recombinantes etiquetadas preferentemente con dominios de unión a colina.
Arevalo Rodriguez Miguel
Cebolla Ramirez Angel
Maestro Garcia-Donas Beatriz
Sanz Morales Jesus Miguel
Velasco Umpierrez Isabel
Biomedal S.l.
Marks & Clerk
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