Chromatography

A well-designed multidimensional (Multi-D) chromatography process, which utilizes optimized individual column methods, can help automate multistep protein purification processes. This can help you save time and increase productivity. See how you can use Multi-D chromatography to expedite the purification of polyhistidine-tagged proteins when using a typical capture, intermediate, and polish purification workflow.

A biomolecule protein purification workflow requires more than one resin. The trick is to find the combination of resins that provides the highest purity in the least number of steps and at the lowest cost. Here are some guides to select the optimal resins for the purification of monoclonal antibodies, native and recombinant proteins, Ig G and mammalian and adenovirus, and for removing aggregates and endotoxins.

Producing highly purified proteins in large quantities is a challenge and requires modification of methods and protocols to attain high efficiency. Dr Murat Kasap from Kocaeli University, Turkey, has overcome these difficulties by taking advantage of the modularity of the NGC Chromatography System and successfully synthesized and purified cardiac troponin-I protein.

Protein purification becomes a challenging step for applications requiring high protein concentrations (such as structural biology studies). This is also true for researchers purifying integral membranes proteins and untagged proteins from native sources. Here we provide some useful tips for selecting the proper resins, optimizing conditions and developing effective strategies to tackle the challenges associated with such difficult-to-purify proteins.

Multicolumn purifications are tedious and the manual operations during this process introduce variabilities, affecting the success of purification and reproducibility of results. This whiteboard animation explains how an automated multi-dimensional chromatography method can make this an easier process with very high success rates.

The separation of monomers from product-related impurities is quite challenging in process separations. It requires a thorough screening and an efficient resin to get efficiency in the purification process. Review the performance data of Bio-Rad’s mixed mode CHT™ Ceramic Hydroxapatite in achieving process separation efficiency and how it compares with various Capto media.

The best chromatography media should yield high protein purity and product recovery as well as be versatile enough to purify difficult samples. Bio-Rad’s CHT™ Ceramic Hydroxyapatite achieves this by virtue of being able to interact with the proteins through multiple modes, making the process extremely efficient. The whiteboard animation describes the mode of action of CHT.

What can chromatography peaks tell us? From the quality of column packing to the purity of separated proteins, a lot can be gleaned from the peaks. Here are some definitions that can help decipher what chromatography peaks can tell us.

Some of the frustrations in protein purification stem from unmet chromatography needs, such as automation, productivity, throughput, and reproducibility among others. Bio-Rad’s new Multi-D chromatography method addresses these and provides solutions to faster, efficient and more reliable protein purification.

Traditional sequential chromatography typically requires intensive hands-on user manipulation, which could hinder the reproducibility of protein purification runs. Multi-D chromatography aims to improve this process while maintaining the efficacy of the traditional workflow.