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Techniques & Tools Sample Preparation, Spectroscopy, Mass Spectrometry, Sensors, Technology, Data Analysis, Food, Beverage & Agriculture, Pharma & Biopharma, Gas Chromatography, Preparative/Process Chromatography

Case study of a semi-pure β-amylase extract

sponsored by Malvern Panalytical

Introduction

Whether you are isolating novel proteins for the first time, investigating the aggregation behavior of purified proteins in different conditions or evaluating the success of your conjugation; ultimately, you have one thing in common: mixtures! And it is in these situations that you will inevitably be reaching for your chromatography columns. Here, we demonstrate how, in the time taken to run a single chromatogram, significant insights into the composition of sample mixtures can be gained from multi-detection size exclusion chromatography (SEC).

Multi-detection SEC combines column chromatography with viscometer, refractive index (RI), ultra violet (UV) and light scattering detectors to provide a powerful analytical tool that can be incorporated simply and effectively into the workflow of those interested in the analysis of protein mixtures.

In this application note, we describe the analysis of a semi-purified protein mixture containing β-amylase (Figure 1). Step-by-step, we show how different properties of the proteins elicit a specific response in each detector and how the coupling this data can be qualitatively and quantitatively evaluated.

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