chromatography

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.

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.

Dr. Mark Snyder, a leading expert in the field of protein purification, presents the purification process development for two very different targets, adenovirus and bacterial surface proteins. In both cases, a two-step column process results in a preparation with high yield and activity and very low host cell protein and DNA contamination. Conditions used make the methods suitable for large-scale production. In this webinar you will learn about: Advantages of mixed-mode resins Optimizing purification process development Procedures for purifying adenovirus and bacterial surface proteins  

Dr. John Bruning (University of Adelaide) and Dr. Jiansheng Wu (Genentech), two leading experts in the field of protein purification, share exciting examples of their research and recent chromatography tools they have used to demonstrate robust and improved efficiencies to their discovery and characterization protein purification workflows.