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Protein purification and bioprocessing

Miniature and parallel

Time to market is crucial in process development, and many companies developing and manufacturing therapeutic agents have invested in automated protein purification and high throughput bioprocessing. The miniaturization and parallelization capabilities of Tecan’s liquid handling platforms ensure an excellent understanding of bioprocesses for robust scale-up into manufacturing, enabling cost-effective development.

Overview

When you need to increase throughput, miniaturized column chromatography is the method of choice.

Key benefits

  • Reduce sample consumption through miniaturization
  • Increase walkaway times
  • Ensure reproducible data for every single run

Chromatography in columns

Make your chromatography processes reliable and time efficient, from simple protein purifications to high throughput process development, bioreactor monitoring and condition screening.

The Freedom EVO® liquid handling platform offers fully automated parallel column chromatography using Repligen’s OPUS® RoboColumn® system, completely eliminating manual processing steps.

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Easy process set-up

The unique Te-Chrom™ Wizard guides the user through the chromatography set-up process step-by-step, generating ready-to-run protocols.

What can it be used for?

  • Condition screening
  • Resin screening
  • Protein purification/separation/concentration
  • Gradient elution
  • Bioreactor monitoring

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Create eight chromatograms in parallel

Column chromatography on the Freedom EVO allows fraction collection down to 25 µl, enabling the creation of chromatograms that match data from complex LC systems. Graphic shows the chromatograms for the separation of two proteins on cation exchangers.

Increase throughput and save valuable time

Parallelization can significantly reduce hands-on time. The Freedom EVO enables fully automated parallel processing of up to eight samples using RoboColumns in just 8 hours, compared to 25 hours using the ÄKTA protein purification system.

For more details see Application Note Pseudo-linear gradient elution

Scalable to your needs

  • Looking for a simple, single-task benchtop solution? Use a Freedom EVO 100
  • Want efficient throughput, increased walkaway times or integrated data analysis? Combine a multimode reader with a Freedom EVO 150
  • Need to automate multiple assays, combining column chromatography and plate-based process development with downstream experiments? Integrate readers, centrifuges and other modules onto a Freedom EVO 200.

Video

Automation of Repligen OPUS®
RoboColumns® on the Freedom EVO

Efficient high throughput process development

Systematic approaches to condition screening shorten the time required to find optimal purification conditions. Automation of PreDictor™ 96-well filter plates prefilled with BioProcess™ chromatography media (GE Healthcare) generates consistent and highly reproducible data.

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Highlights

  • Sophisticated screening for chromatography media and conditions
  • Increased information available
  • Data correlates very well with chromatography column data
  • Can be used with fixed- and disposable tips

Highly reproducible

Capacities for chymotrypsin in a 2 μl PreDictor Capto S plate after 60 min incubation are all within +/-2 standard deviations from the mean.

Reduced sample and resin consumption

Comparison of time and sample consumption using PreDictor plates or true chromatography (Tricorn).

Free-up hands-on time

Comparison of manual (B) versus automated (A) processing of PreDictor plates for both preparation time (yellow) and hands-on time (red).

Affinity purification in plates

Protein purification is a pretty straightforward process, but becomes tedious very quickly when multiple samples have to be handled in parallel. MultiTrap™ 96-well plates from GE Healthcare have been developed to streamline screening of antibodies and tagged proteins.

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What can it be used for?

  • Expression and condition screening
  • Optimization of enrichment conditions
  • Pull-down assays using immunoaffinity techniques
  • Desalting and buffer exchange

Highlights

  • Fully automatable on a liquid handling platform using either vacuum or centrifugation
  • Time efficient workflow that leads to less protein degradation
  • Direct loading, without sample preparation
  • High reproducibility
  • Easy scale-up to ÄKTAxpress™ system (GE Healthcare)
  • Can be used with fixed or disposable tips

Low risk of cross-contamination

SDS-PAGE analysis of clarified E-coli lysates containing two His-tagged proteins (E and K), applied in a ckeckerboard pattern across a His MultiTrap HP plate shows no cross-contamination.

Chromatography in tips

What if purifying 96 protein samples only took 15 minutes with minimal hands-on time?

PhyTip® columns are designed for use with liquid handling platforms equipped with 8- or 96-channel pipetting arms, offering walkaway protein purification for higher throughput demands.

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Highlights

  • Purification of native or recombinant proteins or analytes
  • Minimal cross-contamination
  • High yields, with elution volumes down to 10 μl
  • Packable with standard or custom resins
  • Usable with disposable tips only

High reproducibility

The Freedom EVO offers precise and accurate liquid handling using PhyTips, ensuring highly reproducible results (SD = 0.5)

Exceptional mass recovery

Immunoglobolin G sample volumes of 200 or 500 ul were loaded onto 200+ and 1000+ PhyTips Protein A columns respectively. The mass recovery of IgG as a function of column bed size is shown. The measured binding capacity of the column is 29 µg/µl bed volume.

Data kindly provided by PhyNexus

Protein affinity capture tips

Benefit from a one-stop solution for affinity purification of large molecules with Thermo Fisher Scientific’s MSIA™ (Mass Spectrometric Immunoassay) Streptavidin-EVO microcolumns.

MSIA Streptavidin affinity capture tips are affinity microcolumns with the ability to capture and purify antibodies and other proteins from various samples.

Overview

Our systems are scalable and can be tailored to your individual needs. From simple benchtop solutions to complex systems that run multiple assays on a single instrument, we can provide laboratory automation to scale up your processes.

Key benefits

  • Miniaturization reduces sample, reagent and resin consumption, saving money
  • Template scripts available

Keep it simple

PARALLEL ANTIBODY PURIFICATION

The easiest way to improve data reproducibility and save hands-on time is to automate Repligen’s proven RoboColumn® technology on the Freedom EVO®. Equipped with an eight-channel liquid displacement pipetting arm and the unique Te-Chrom Wizard, the Freedom EVO parallel antibody purification workstation allows fast, comprehensive set-up of all processing steps. Affinity chromatography purified antibodies for downstream mass spectrometry or binding studies are ready in short time.

Highlights

  • Purify antibodies for a variety of downstream applications including ELISA, western blotting, immunohistochemistry, immunoprecipitation and LC-MS
  • Run 96 samples in 5.6 hours – freeing your time for other work
  • Process eight analysis-ready samples in 30 minutes
  • Small footprint, only 1.08 x 0.78 m

Example workflow showing automated (blue) and manual (gray) process steps.

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Add fully automated plate handling

SIMULTANEOUS STEP-GRADIENT ELUTION FOR TWO PROTEINS

Add a Robotic Manipulator Arm to the Freedom EVO 150 platform to provide automated plate handling for additional walkaway time. This platform comes with an eight-channel liquid displacement pipetting arm and a Te-Chrom™ Shuttle that enables the collection of fractional eluates using RoboColumns.

Highlights

  • Additional space allows you to automate additional workflows
  • Compact footprint, 1.45 x 0.78 m
  • Te-Chrom Wizard for fast and comprehensive process set-up, from column equilibration to fraction collection
  • Buffer Creation Wizard – prepare 96 individual buffers in 10 minutes
  • Process eight samples and collect six fractions from each in just 16 minutes
  • Reproducible data comparable to LC column systems

Example workflow showing automated (blue) and manual (gray) process steps.

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Add data collection

PARALLEL CHROMATOGRAPHY AND DATA COLLECTION

Automation not only ensures reproducible data and frees up hands-on time, it also helps to reduce costs by minimizing consumption of precious samples, reagents and resins. The development time for new target molecules can be noticeably shortened by parallelization of up to eight chromatographic separations, with subsequent analysis using an integrated microplate reader.

Highlights

  • Full automation, including plate handling and data collection
  • Proven liquid displacement pipetting technology
  • Freedom EVO 150 with a compact footprint of 1.45 x 0.78m
  • No need for programming expertise – set up your instrument with the help of a graphical wizard
  • Set up 96 individual buffers in just 10 minutes – let the Buffer Creation Wizard calculate all the required volumes.

Example workflow with all processing steps fully automated.

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Overview

Discover our software solutions to easily set up complex protein purification processes.

Key benefits

  • No need for programming expertise – set up your protocol with the help of a graphical wizard
  • Define your buffers – let us calculate and pipette up to 96 different conditions
  • From stock solutions to ready-to-use buffers in a few clicks

Column chromatography set-up

Setting up chromatography processes can be complex, making the programming of automated systems difficult. With the unique Te-Chrom Wizard, we have made programming fast, simple and reusable.

Creating a ready-to-run Freedom EVOware® scripts is now a matter of a few clicks. Easily set up all the processing steps – from column equilibration to sample addition and elution including fraction collection and detection. Data can then be used to generate chromatograms and full traceability is guaranteed using the system’s Sample Tracking feature.

Highlights

  • Extended residence time due to slow flow rates
  • Automated plate logistics and data collection
  • Neutralization of purified sample
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Buffer preparation

The number of different buffer compositions required for protein purification condition screening or high throughput process development can quickly become major headache. The Buffer Creation Wizard add-on for Freedom EVOware® allows fully automated preparation of buffer sets for a wide range of applications.

The Buffer Creation Wizard guides the user through six steps to input all the information necessary for the process, including stock solutions, destination plate or tube format, etc. The wizard then automatically calculates the required volumes of each stock solution for every well or tube, and provides a ‘shopping list’ of all required stock solutions to simplify worktable set-up. Finally, it generates a ready-to-run Freedom EVOware script which can be saved and run at any time.

Highlights

  • Easy to operate graphical user interface
  • Customizable stock solution and buffer libraries
  • Straightforward plate layout creation in the wizard, or easy import from DOE software, such as Assist (GE Healthcare), MODDE® (Umetrics) or JMP® (SAS)
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Videos

Tecan Buffer Creation Wizard

Tecan Chromatograpy Wizard

Brochures

The Freedom EVO platform offers high throughput automated protein purification and bioprocessing for the development and manufacture of therapeutic agents. The miniaturization and parallelization capabilities of the platform ensure a robust scale-up process, guaranteeing cost-and time-efficient development.

Using the OPUS® RoboColumns on the Freedom EVO workstation enables true parallel chromatography of up to eight columns, offering rapid screening of a range of parameters in the shortest time frame possible, without using sophisticated LC systems.

Application Notes

This application note describes how MSD is using RoboColumns® on the Freedom EVO® platform to qualify HTPD methods for the characterization of mAb purification steps, for both bind and-elute cation exchange (CEX) and flow-through anion exchange (AEX) studies.

Purification of biopharmaceuticals is still dominated by sequential liquid chromatography in a packed column bed, which is not only difficult and time consuming, but also very expensive. This application note describes the implementation of automated resin screening using Repligen’s OPUS RoboColumns on the Freedom EVO liquid handling platform.

Combining PhyTip columns with the Freedom EVO platform offers high throughput automation for sample preparation and protein purification.

The Freedom EVO platform offers complete automation of sample preparation and protein purification using PhyTip columns, allowing straightforward process development and predictable scale-up procedures.

The Freedom EVO workstation can be used to screen chromatographic process parameters, helping to accelerate development of biopharmaceutical purification processes. The system can be used with a wide range of modules and accessories, such as the MediaScout® RoboColumns® miniaturized chromatography columns.

Posters

Automated process development on the Freedom EVO platform can accelerate development of both upstream and downstream processes for large-scale bioproduction, as well as improving process analytics.

Automation with the Freedom EVO to accelerate cell culture upscaling, downstream process development and process analytical technology (PAT).

An overview of chromatographic process development tools on the Freedom EVO platform, including finite bath thermodynamic studies, batch binding in PreDictor™ plates, bind/elution studies, and breakthrough/elution experiments with MediaScout RoboColumns.

Tecan Journal Articles

Biomanufacturing requires careful separation of the molecule of interest from other cellular products to ensure the quality and stability of the final product. This is even more important for the manufacture of biotherapeutics, as the presence of unwanted molecules in pharmaceutical preparations can affect the efficacy and safety of biologically-derived drugs. FUJIFILM Diosynth Biotechnologies is using automated chromatography condition screening to generate more data in less time, helping to improve the performance of purification processes and accelerate biological drug manufacturing.

The biosimilars market is expanding rapidly as the patents expire for an increasing number of high profile biopharmaceutical agents. The complex nature of biologics requires extensive characterization of the production techniques and in vivo effects of new biosimilars before they can be released onto the market. Coherus Biosciences is using advanced laboratory automation to help screen chromatography conditions as part of its downstream purification processes for new biosimilars.

Merck KGaA has implemented automated miniature column chromatography protocols to improve throughput and reduce the amount of material required for the development and characterization of new chromatography media, allowing more measurements to be performed in less time.

SomaLogic's SOMAscan® proteomics technology is helping to identify new biomarkers for a wide range of disease states. Capable of simultaneously measuring thousands of proteins in low volume biological samples, this technique is ideal for the detection of low abundance proteins.

Thermo Fisher Scientific has developed innovative sample processing tools designed to simplify and accelerate analytical workflows. Offering automation-friendly affinity purification for proteins and antibodies, this provides a robust solution for high throughput preclinical research and development.

UCL’s Advanced Centre for Biochemical Engineering is harnessing the flexible automation capabilities of its Freedom EVO workstations to explore novel bioprocessing applications for the production of ‘next generation’ medicines, including protein-derived therapeutics, vaccines and cells for therapy.

Tecan and GE Healthcare are working together to increase the quality and consistency of high throughput protein purification for process development, combining the Freedom EVO workstation with PreDictor 96-well plates pre-filled with BioProcess™ chromatography media.

UCB has increased the throughput of its antibody purification pipeline by using a Freedom EVO workstation to miniaturize its chromatography applications, allowing the company to investigate far more parameters in a fraction of the time, while consuming significantly less material.

The combination of the Freedom EVO platform and Atoll’s MediaScout RoboColumns offers a powerful tool for fully automated high throughput process development, allowing valid chromatographic conditions to be used without the need for sophisticated LC systems.

Scientists at Bayer Schering Pharma’s Berlin research facility have integrated two Freedom EVO workstations into their protein production process, including automation of GE Healthcare’s MultiTrap® 96-well plates.

For research use only.