chromatography

When testing new drug therapies, it is essential to know the identity of the protein being tested. Chromatography is a commonly used technique to separate complex biological mixtures and isolate proteins of interest prior to downstream characterization. Learn more about how automating chromatography and stain-free technology are helping to save time and effort in protein purification.

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.

We’ve combined the chromatography process with purity and yield assessment using stain-free gels to shorten and optimize your protein purification workflow. See how it’s done.

This video describes the three main steps in attaching a DynaLoop Sample Loop to the NGC™ Chromatography System to enable the injection of larger sample volumes: connecting the DynaLoop Sample Loop to the NGC Chromatography System, priming the DynaLoop Sample Loop, and creating a method in ChromLab™ Software.

The NGC pH valve provides real-time, in-line monitoring of buffer pH. In this video, you’ll learn how to connect the pH valve to the NGC System, add it to the flow path, and calibrate the valve using buffers at two different pH values. Features of the pH valve include accessible calibration ports and easy bypass of the probe when the NGC System is being cleaned.

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.

This video explains how to remove bubbles and/or reduce bubble formation in your NGC System. Tips to reduce bubbles include warming up your multiwave detector for 30 to 60 min before use and flowing buffer at 0.1–0.2 ml/min in the manual mode until a stable/flat baseline is achieved before starting a method. The system may also be flushed with 100% methanol or ethanol at purge speed, with columns out of line, to remove bubbles.

This video describes the functions of the Air Sensor Module and how to connect it to the NGC Chromatography System. The NGC Air Sensor senses the end of sample or buffer and prevents the introduction of air into the system, which can damage columns and/or incur the loss of precious samples. Both small and large air sensors are available to fit the tubing being used. Steps for connecting the Air Sensor Module and associated air sensors are described including how to add the air sensors into the system flow path using ChromLab™ Software and the differences in using the air sensors in manual vs. method modes.

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.