Laboratory Products

Driving Productivity in Protein Purification

Nov 04 2014

Author: Per Lidén (GE Healthcare Life Sciences), Svend Kjær (Cancer Research UK) and Mark Cooper (Abcam) on behalf of GE Healthcare

Free to read

This article has been unlocked and is ready to read.

Download

Crucial to the successful operation of any research laboratory is the ability to efficiently generate purified proteins for further investigation and analysis. In addition to timely delivery, researchers also demand a protein product which is pure, stable and active – it is crucial for antibodies to have the necessary binding activity and enzymes must be fully functional.
How a laboratory, or indeed a company or institution as a whole, goes about addressing these needs, can vary significantly and there are a range of technologies, workflows and organisational structures that can be implemented.

Challenges to Productivity
Identifying and understanding current bottlenecks and challenges to protein purification is the first step towards improving productivity to deliver positive business benefits.
As a global supplier of specialist protein research tools, Abcam intimately understands these challenges as it works to provide large numbers of high-quality complex protein-based products to researchers with varying requirements. The company utilises a number of chromatography systems for protein purification across multiple sites globally. Increasing demand globally, created a need to increase production of small volumes of high-quality proteins at high throughput consistently, irrespective of geographic location, from its existing workforce. This challenge was coupled with the requirement to maintain levels of expertise and technical support, thereby potentially increasing pressure on in-house experts.
Similarly, the Protein Purification Facility (PPF) at the London Research Institute (LRI), Cancer Research UK (CRUK) expresses and purifies recombinant proteins for applications with high quality demands such as x-ray crystallography and downstream analysis of functional activity e.g. binding and/or enzymatic activity. Critical to success is minimising hands-on interaction with the proteins, many of which are highly sensitive, to preserve enzymatic activity. As a consequence, speed and access to instruments are key considerations. An internal review of all operations across the PPF identified that the existing batch-based purification approach, involving significant amounts of hands-on work, could be targeted for streamlining, to increase output and improve throughput. 

Practical Approaches to Improving Productivity
Once the bottlenecks in a protein purification workflow have been identified and understood, it is necessary to evaluate options for addressing them.  
Productivity in protein purification can be improved through use of automation that reduces hands-on technical requirements, reducing the chances of error and streamlining production efficiency and work effort. Simplification in the use of equipment can support fast set-up of protocols and improve utilisation while maintaining flexibility and the ability to carry out complex procedures. Standardised approaches, in addition to improved system design, reduces training requirements, boosting productivity for experienced scientists while opening up protein purification technologies to researchers who are not necessarily protein purification experts. As resource utilisation is improved the overall output from available instruments can increase significantly, ultimately leading to fewer instruments required, which in turn means savings on maintenance work, service support contracts and lab space.

Globally Distributed Manufacturing Delivered through Standardisation
The purchase of multiple automated protein purification systems by Abcam has further enhanced both productivity and quality, and has been used to support its latest innovations, specifically, the manufacture of its Alexa Fluor®-conjugated secondary antibody product range. Innovation, in-house manufacturing, and quality are significant factors in the success of Abcam today, and the decision to automate over traditional hands-on approaches was to ensure quality of the final product (monitoring & control of process parameters), in addition to increased productivity/throughput. After the Alexa Fluor®-conjugated secondary antibody range was launched in December 2012 production batch sizes needed to scale up 5-fold to meet sales demand and generate sufficient stock. ÄKTA™ pure (from GE Healthcare) was flexible enough to allow scale-up of production, whilst maintaining necessary throughput.
Having a global customer base meant that global standardisation was important, as was security of supply. A combination of increased in-house production, as well as developing relationships with multiple suppliers, enabled Abcam to maintain provision of its large catalogue of protein research tools without any reduction in levels of expertise and technical support. Platforms such as the ÄKTA protein purification systems permit the production of the same protein to a consistent standard at any of Abcam’s international sites. This arrangement allows faster restock of products to maintain customer supply despite the increased demand. The interoperability that exists across the ÄKTA systems range allows Abcam’s protein purification scientists to move easily between different system sizes in order to maximise cost-effectiveness.

Purification Time Halved through Automated Workflow
By moving to automated protein purification systems (ÄKTA pure from GE Healthcare) at the PPF at CRUK, the continuous running of the instruments is now possible. For example, a batch of labile GST-tagged kinases would previously take 3-4 working days but can now be left overnight to run so it is purified within 2 working days. The increased flexibility of the automated workflow means that, at the initial stages of developing a protocol, challenges in purification can be overcome creatively. Then, once a good protocol has been achieved, it can be formalised for simple re-use. For example, proteins that precipitate quickly after being eluted from a column, especially when left to run overnight, can now be directly loaded into a buffer exchange column.  In addition, the increase in automation eliminated a manual step, meaning less manual protein handling and resulting in higher protein quality and yields.

Save Scientists’ Time Through Intuitive Software
Methods for ÄKTA protein purification systems are set up through the ‘drag and drop’ interface of the UNICORN™ control software, which has enabled scientists at Abcam and CRUK to put together easy-to-use, intuitive protocols quickly, and also reduced the training need. For Abcam, movement from development into production, whilst maintaining flexibility, is supported by the fact that this easy-to-program software is installed platform-wide. For CRUK, the fact that many of the columns it uses are pre-programed into the software saves further time.

Conclusions
At Abcam, use of automated protein purification platforms such as ÄKTA pure has helped to achieve improved process economy, while reducing the length of training and allowing innovative processes to be set up quickly. Use of an easy-to-use, automated system for protein purification has resulted in significant improvements to team productivity while facilitating the global uptake of innovative methods developed in-house.
At CRUK, the introduction of an automated protein purification platform was able to more than halve the time for a typical protein purification run, whilst maintaining the flexibility required for the set-up of more challenging proteins.
Addressing bottlenecks in protein production and purification with the application of controlled, automated and standardised purification platforms is a key step to driving innovation. Straightforward and easy-to-learn workflows allow for quick programming of protocols and sharing across multiple sites, enabling users to achieve a standardised level of protein purification, and resulting in efficient production of pure and functional proteins.

Alexa Fluor® is a registered trademark of Life Technologies. Alexa Fluor® dye conjugates contain(s) technology licensed to Abcam by Life Technologies.
ÄKTA™ and UNICORN™ are trademarks of GE Healthcare companies.

Free to read

This article has been unlocked and is ready to read.

Download


Digital Edition

Labmate UK & Ireland 49.2 - March 2024

March 2024

In This Edition Articles - Choosing the right LIMS for your lab: Four key considerations - Pay increases to continue but staff focus on security and training - Building a global culture of...

View all digital editions

Events

Making Pharmaceuticals

Mar 26 2024 Milan, Italy

FORUM LABO LYON

Mar 27 2024 Lyon, France

Microbiology Society Annual Conference 2024

Apr 08 2024 Edinburgh 2024

analytica 2024

Apr 09 2024 Munich, Germany

ChemBio Finland 2024

Apr 10 2024 Helsinki, Finland

View all events