Many laboratories today are faced with producing timely and accurate results at reduced cost. The pace of life science R&D and manufacturing is being accelerated but the hours in a single day has not changed. The need for companies to shorten their development or manufacturing time means that process development must be fast and inexpensive without compromising the quality of the end product. To address this challenge, many companies are moving towards automation. This trend is definitely being seen in the chromatography space with the advent of multimodal chromatography process improvement and new multidimensional (multi-D) chromatography instruments. Although this concept has been in practice for decades, the instrumentation and software technology for multi-D chromatography continues to evolve – here we discuss what all the excitement is about.
What is multi-D chromatography
Multi-D chromatography is a powerful technique for separating complex and difficult substances. There are a lot of highly complex mixtures, which researchers would like to separate for qualitative and quantitative analysis, but due to their complex make-up, this can be very difficult to perform. Highly complex mixtures can be separated by passing the sample through two or more separation stages. In a multi-D workflow, the eluate from the initial column flows past a detector and is transported into a secondary storage before being applied to the next column. This enables researchers to separate compounds using multiple properties and potentially generating greater selected end-product. The more properties used, the cleaner the resulting sample. Standard analysis techniques such as SDS-PAGE or analytical size-exclusion chromatography (SEC) can be used to analyze sample material after a multi-D run. Traditionally, this workflow requires that the user be present to manipulate columns and collectors throughout the long purification. This is a very time-consuming and potentially error-prone process. Today, instruments and software packages have automated this powerful continuous multi-step purification allowing the user to walk away and focus on other work but still have the confidence that they will end up with a high quality purified sample.
Why Is automated multi-D so great?
Convenience
In the past, researchers ran instruments in manual mode due to the complexity of their purification procedures. Those days of manually collecting fractions and moving your sample from one column and injecting onto another are a thing of the past. Software packages today introduce template workflows that cut through the busy work of protein purification. These technology advances transform an extensive hands-on purification process to ‘touch button’ purification. Many systems on the market are easy to operate with everything controlled via push buttons and easy-to-navigate displays and software. Guides have been created to help setup and run tandem or multi-D purification workflow. The system software takes in to consideration several upfront factors and variables in order to make creating and validating new workflows easier. For added convenience, many vendors have introduced pre-programmed templates and methods all optimized for defined columns. Parameters are preset so all you need to do is enter sample volume and press “Go”.
Reproducibility and Reliability
Walk-away convenience isn’t the only advantage – human induced variability, which is nearly impossible to avoid when dealing with any chromatography instrument directly, is taken away. Traditional sequential chromatography typically requires intensive hands-on user manipulation that could hamper the reproducibility of protein purification runs. Because people aren’t machines, there is greater variability from experiment to experiment introduced through pipetting errors, tubes spilling or sample mixed-up. Automating protein purification ensures more reliable results. With instrument and software packages such as Bio-Rad’s automated NGC System and ChromLab Software, you can eliminate the problem of sample loss through spillage or by misplacing tubes by letting the instrument do the work for you. And because the system performs your program consistently, you can be assured of reliably high purity levels and consistent reproducibility.
Time to results
There never seems to be enough time in a day to do everything that needs to be done. Life science research, product development and manufacturing are being pushed to do more with less resources and time. The last thing managers want to see is their workforce being consumed by a lengthy hands-on procedure when they could be planning experiments, analyzing results or writing papers. With the introduction of multi-D chromatography instruments and software, complex purifications that could take days to do can now be done in a single day or even hours giving researcher time to do other tasks.
Addressing unmet needs
To sum it all up, with multi-D workflow templates and automation, there is no need for direct and constant manual interaction. The hands-on time drops dramatically while purification levels and reproducibility escalates. No more baby-sitting your experiment with these new systems. They save you time and headache while increasing the reproducibility and purity of your protein purification. So go ahead start your purification and walk away. Don’t you have something better to do?
Image: Courtesy of Bio-Rad, Multi-D workflow representation
Related Products from: Bio-Rad