Fast Screening for the Purification of Proteins Using Membrane Adsorber Technology

Abstract

New membrane systems for the purification of proteins are presented. These disposable devices enable time saving screenings of downstream conditions with a small amount of protein solution. Furthermore, scaling up to the production level is directly possible since devices with effective membrane surfaces from 10 cm2 to 1 m2 are available. Several membranes like ion exchange membranes were investigated for the determination of protein binding capacities. Metal chelate membranes were used for ligand binding, which enables the determination of the membrane systems potential for affinity chromatography. Lysozyme and BSA are chosen for the determination of binding capacities of ion exchange membranes. Furthermore, Con A membranes, suitable for the isolation of glycoproteins, were developed and their binding properties were tested.

Automated Microscale His‐tagged Protein Purification Using Ni‐NTA Magnetic Agarose Beads

“…Protein purification is an important step in many biological experimental procedures such as protein function analysis [1] and protein screening research [2,3]. The development of biotechnology and proteomics involves the purification of a single protein or multiple proteins from hundreds of microscaled bacterial culture samples [4].…”

One-step-purification of penicillin G amidase from cell lysate using ion-exchange membrane adsorbers

“…First the so called "negative" separation of the enzyme by depletion of all host cell proteins through binding them unspecifically to an ion exchange material, thus collecting the PGA in the flow-through [25] and second the specific binding of PGA itself to an ion exchange material, keeping all host cell proteins (HCP) in the flow-through and subsequent elution of the bound enzyme [9]. Both strategies were tested in the described microliter scale highthroughput screening (see chapter 2.2) with cell lysate to identify best binding material and evaluate basic process conditions.…”

“…Many papers deal with the use of membrane adsorption for purification and isolation of proteins [26][27][28] and antibodies or to remove DNA, host cell proteins, endotoxins und viruses respectively [2,[7][8][9][10][11][12][13][14]. Membrane adsorbers employ the same strong anion and cation exchange ligands, quarternary ammonium (Q) respectively sulfonic acid (S), as standard ion exchange chromatography columns.…”

Considerations on the flow configuration of membrane chromatography devices for the purification of human basic fibroblast growth factor from crude lysates

“…Membrane chromatography offers many advantages for the design of cost‐effective downstream processing . It is frequently used for polishing applications in biotechnological processes for the fast processing of large volumes of solutions.…”