Lab-on-a-Tip: Reversible adsortive foams for protein and DNA purification

MIRNA SANCHEZ; MARCELO FERNANDEZ LAHORE; MARIANO GRASSELLI

Beijing

Congreso; 17th International Biophysics Congress –The 12 nd National Biophysics Congress; 2011

Biophysical Society of China

Modern biotechnology heavily depends on the availability of efficient processes, which should be able to generate competitive products. Recovery and purification of bioproducts is mainly based on chromatographic separations. However, the low productivity imposed by diffusional limitations, the need for multiple chromatographic steps, and the requirement of extensive solid separation, drives to a technological situation characterised by low global product yields. Disposable products reduce the process development and costly validation protocols. Our strategy is combine direct product separation and disposable materials. The aim of this work is to develop novel chromatographic supports based on synthetic foams for protein capture from non-clarified feedstocks. Polyurethane foams were coated with poly(glycidylmethacrylate) (pGMA) by radiation-induced polymerization. The grafted reactive polymer is further modified with sulfonic acid, iminodiacetic acid (IDA) or silica dioxide in a second step. Adsortive foams were packed in a micropipette tip and used for recovery lysozyme (by cation-exchange process) or recombinant His-tag green fluorescent protein (GFP-His) from crude protein mixtures (by IMAC process). DNA purification was performed by tips modified with silica dioxide. Polymeric foams are promising chromatographic matrices for recovery and purification of biomolecules