Recombinant Protein Purification

Recombinant protein purification is the process of isolating genetically engineered proteins expressed in host systems such as E. coli, yeast, or mammalian systems. These proteins are often produced with affinity tags that enable selective purification and improve recovery across a range of purification methods. The objective is to obtain a pure, functional protein suitable for downstream applications including structural studies, activity assays, or therapeutic development.

We offer a comprehensive portfolio of magnetic beads and chromatography resins designed to support recombinant protein purification across multiple expression systems and purification workflows. Our affinity purification solutions are available in scalable formats to meet needs of high-throughput screening, routine laboratory purification, and pilot- to process-scale applications.

• Broad tag compatibility: Affinity supports are available for purification of recombinant proteins containing common fusion tags, including 6xHis, GST, DYKDDDDK (FLAG), c-Myc, and HA.
• Flexible formats and scalability: Magnetic beads, magnetic agarose, loose resins, spin columns and kits, FPLC cartridges, and 96-well spin plates support purification from microgram- to low-kilogram-scale workflows.
• High-performance purification: Optimized resin chemistries help deliver high binding capacity, strong recovery, and low nonspecific binding across diverse protein targets.

Producing purified recombinant protein involves a series of coordinated steps, from gene design and cloning to expression, harvesting, and downstream purification. Below is an overview of the key steps typically followed to obtain purified recombinant proteins.

Figure 1. High-yield, high-purity, medium-scale purification of 6xHis-Tagged protein. More than 4 grams of over-expressed 6xHis-GFP were purified in 3 hours using 200 mL columns containing HisPur Ni-NTA Superflow Agarose or Qiagen Ni-NTA Superflow. One liter of lysate was loaded at a flow rate of 20 mL/min, then washed until baseline with wash buffer containing 30 mM imidazole. Bound protein was eluted with buffer containing 300 mM imidazole. Fractions containing purified 6xHis-GFP were pooled and quantitated using Pierce 660 nm Protein Assay. Load, flow-through, wash, and eluate fractions were separated by SDS-PAGE, stained with Imperial Protein Stain and evaluated using myImageAnalysis Software to determine purity.

See experiment details and additional data

Figure 2. Thermo Scientific Pierce Glutathione Agarose facilitates high yield and high purity GST-fusion protein purification. E. coli lysate (14.4 mg total protein) containing overexpressed GST was incubated with 50 µL GSH resin from various suppliers and purified per manufacturers' instructions. The amount of GST eluted from the resin (yield) was quantified by Thermo Scientific Coomassie Plus Protein Assay. Purity was assessed by densitometry of the stained gel lanes. M= MW marker; L=Lysate load; FT=Flow-through; E=Elution.