Introduction Improved purification methods are required for protein structure and function studies, necessitated by growing demands from proteomics, drug development, and biotechnology programs. To simplify purification of recombinant proteins, including many with unknown biochemical properties, several genetically engineered affinity tags, or purification tags, are used. Commonly used tags are...

Previously, we reported a non-chromatographic protein purification method exploiting the highly specific interaction between the dockerin and cohesin domains from Clostridium thermocellum and the reversible aggregation property of elastin-like polypeptide (ELP) to provide fast and cost-effective protein purification. However, the bound dockerin-intein tag cannot be completely dissociated from t...

Affinity purification is based on the selective and reversible interaction between two binding partners, of which one is bound to a chromatography matrix and the other may be either a native target protein or a recombinant protein fused with an affinity tag (Cuatrecasas et al. 1968). Recombinant DNA-technology allows straightforward construction of gene fusions to provide fusion proteins with t...

The fusion protein of green fluorescent protein (GFP) and human interleukin-2 (hIL-2) was produced in insect Trichoplusia ni larvae infected with recombinant baculovirus derived from the Autographa californica nuclear polyhedrosis virus (AcNPV). This fusion protein was composed of a metal ion binding site (His)6 for rapid one-step purification using immobilized metal affinity chromatography (IM...

BACKGROUND Recombinant protein purification is a crucial step for biochemistry and structural biology fields. Rapid robust purification methods utilize various peptide or protein tags fused to the target protein for affinity purification using corresponding matrices and to enhance solubility. However, affinity/solubility-tags often need to be removed in order to conduct functional and structura...

Recent advances in genomics, proteomics, and bioinformatics have facilitated the use of recombinant DNA technology in order to evaluate any protein of interest, without prior knowledge of the protein’s cellular location or function. The parallel use of affinity tags with recombinant DNA techniques, allows the facile modification of proteins of interest leading to efficient identification, produ...

Cancer therapy with oncolytic measles virus (MV) requires high titers of pure, infectious particles (~108 times more per dose than the vaccine). A gentle but efficient downstream purification process is therefore essential. Here we investigated suitability ion-exchange chromatography for MV using resin-based stationary phases. The success was evaluated based on yield, ratio to total particles, ...

BACKGROUND Hydrophobic interaction chromatography (HIC) most commonly requires experimental determination (i.e., scouting) in order to select an optimal chromatographic medium for purifying a given target protein. Neither a two-step purification of untagged green fluorescent protein (GFP) from crude bacterial lysate using sequential HIC and size exclusion chromatography (SEC), nor HIC column sc...

Functional genomics requires structural and functional studies of a large number of proteins. While the production of proteins through over-expression in cultured cells is a relatively routine procedure, the subsequent protein purification from the cell lysate often represents a significant challenge. The most direct way of protein purification from a cell lysate is affinity purification using ...