MarvoBlog — Insect cell protein purification

     The purification conditions allowing for the maximal protein yield and purity may vary significantly depending on the characteristics of a given target protein. Finding the best conditions frequently require optimizing the key parameters through small-scale trial and error approaches. 1. Setting up binding conditions for target protein      In most successful purification protocols, the ratio for binding is 1 mL of resins for every 10 – 50 mL of eukaryotic culture supernatant; and 1 mL for 10 ~ 20 mL E.coli lysate.      To set up protocol to purify a new protein, it is recommended to start with the...

Our Ni-Penta™ Agarose-Base Resins, new members of our ProteIndex™ family, are advanced, chemical-tolerant immobilized metal ion affinity chromatography (IMAC) media that are precharged with nickel ions. Via a proprietary pentadentate ligand, immobilization of nickel ions on the matrix surface is highly uniform and stable, and offers excellent resistance and anti-strip power to a wide range of chemicals, including NaOH, EDTA, reducing agents DTT and β-ME. Ni-Penta™ resins are well suited for direct capture of polyhistidine-tagged proteins that are secreted into eukaryotic cell culture supernatant (i.e. CHO cells, other mammalian cells, and insect cells). Unlike the conventional IMAC resins, purification with...