Protein purification and bioprocessing MINIATURE PARALLEL AUTOMATED

Transcription

1 Protein purification and bioprocessing MINIATURE PARALLEL AUTOMATED

2 2 Parallel chromatography in columns OPUS ROBOCOLUMNS CHROMATOGRAPHY MINIATURIZED Combining Repligen s miniaturized chromatography columns with Tecan s Freedom EVO or Fluent liquid handling platforms enables fully automated parallel column chromatography. Up to 96 OPUS RoboColumns, pre-packed with chromatography resin, can be individually arranged on a 96-position array to meet specific application requirements. LIQUID HANDLING AND SOPHISTICATED AUTOMATION Tecan s liquid displacement pipetting technology ensures successful processing of up to eight RoboColumns in parallel, with continuous flow dispensing for volumes of up to 4.8 ml per pipetting step. The systems offer a choice of simple batch elution (for sample preparation) or individual fraction collection, enabling the generation of chromatograms that match complex LC system data. This ensures excellent robustness of the scale-up process into manufacture. APPLICATIONS Condition screening to determine optimum purification conditions Resin screening to test different resins in parallel Sample preparation for downstream applications, such as protein crystallography or MS analysis Process analytics for bioreactor monitoring, to determine the optimal harvesting point The chromatogram (right) shows the separation of mg/ml each of lysozyme and cytochrome c on eight OPUS RoboColumns. The columns were packed with cation exchange resin (2 μl Toyopearl SP655) and processed in parallel on a Freedom EVO. Elution was performed using.1 M NaCl for ODl=28nm RoboColumn 1 RoboColumn 2 RoboColumn 3 RoboColumn 4 RoboColumn 5 RoboColumn 6 RoboColumn 7 RoboColumn 8 Elution buffer [%] Elution buffer [%] lysozyme, and 1 M NaCl for cytochrome c (Data kindly provided by Repligen) V [ml]

3 3 Software and solutions PROCESS SET-UP SIMPLIFIED Tecan s solutions offer easy set-up of all processing steps from column equilibration to sample addition and elution including fraction collection and detection. You have a choice of: Freedom EVO platform with Freedom EVOware software offering a unique Te-Chrom Wizard for process set-up, and a Buffer Creation Wizard for fully automated buffer preparation Fluent Automation Workstation with FluentControl software using its powerful and intuitive TouchTools touchscreen interface to simplify day-to-day operation EXPLORE OUR FREEDOM EVO-BASED SOLUTIONS Antibody purification Affinity chromatography-purified antibodies can be quickly prepared for downstream MS analysis or binding studies. Step gradient elution Fully automated plate handling allows simultaneous step-gradient elution, enabling you to collect individual fractions and find the optimal separation conditions for your proteins. Condition screening Increase walkaway time and add automated data collection and analysis to your workflow using an integrated microplate reader.

4 4 Protein purification in plates PREDICTOR BIOPROCESSING A systematic approach shortens the time required to find optimal purification conditions. Automated processing of PreDictor 96-well filter plates prefilled with BioProcess chromatography media (GE Healthcare) generates consistent and highly reproducible data. Reproducibility of chymotrypsin capacities was determined for a 2 μl PreDictor Capto S plate (6 min incubation). The solid red line corresponds to the average value, dashed lines denote ±2 standard deviations, and columns 1 to 12 are denoted by the alternating colored bars Preparation time and hands-on time were compared for the automated process on a Freedom EVO (A) and the manual A process (M). Yellow denotes the preparation time, and red denotes the hands-on time during processing. M (Data kindly provided by GE Healthcare) Time (min) 12 Preparation time Hands on time during process MULTITRAP AFFINITY PURIFICATION IN PLATES Protein purification is a reasonably straightforward process, but becomes tedious very quickly when multiple samples have to be handled in parallel. MultiTrap 96-well plates (GE Healthcare) have been developed to streamline screening of antibodies and tagged proteins. C l a r i fi e de. coli lysates containing two His-tagged proteins (E and K) were applied in a checkerboard pattern across a His MultiTrap HP plate, then analyzed by SDS-PAGE. Results indicated no cross-contamination. (Data kindly provided by GE Healthcare)

5 5 Protein purification in tips PHYTIP COLUMNS PURIFICATION IN A PIPETTE TIP Purification of analytes, or native or recombinant proteins, can be performed in as little as 15 minutes with PhyTip columns. Designed for use with liquid handling platforms equipped with eight- or 96-channel pipetting arms, they offer walkaway protein purification for higher throughput demands. Sample volumes of 2 or 5 μl Immunoglobulin G (IgG) were loaded onto 2+ and 1+ PhyTip Protein A columns respectively, then the mass of IgG recovered was calculated as a function of column bed size. This indicated a binding capacity of 29 μg per μl bed volume. (Data kindly provided by PhyNexus) Mass Recovery (µg) Resin Bed Volume (µl) MSIA TIPS AFFINITY PURIFICATION FOR MASS SPECTROMETRY Thermo Fisher Scientific s MSIA (Mass Spectrometric Immunoassay) Streptavidin-EVO microcolumns offer a one-step solution for affinity purification of antibodies and other proteins. Save time by processing 96 samples in parallel. Ligand Binding-MSIA (LB-MSIA) workflow was performed on a 2 μl rodent plasma sample containing 1 μg/ml of adalimumab and 35 μg/ml SILu Lite. The deconvolved spectrum shows the masses and peak intensities for SILuLite (IS) (146,824 Da) and adalimumab (148,81 Da). (Data kindly provided by Thermo Fisher Scientific)

6 Protein purification. Bioprocessing. Process analytical technology Discover what Tecan offers for your chromatography needs Contact your local Tecan representative to define the chromatography workstation that fits your needs. For research use only not for use in diagnostic procedures. Australia Austria Belgium China France Germany Italy Japan Netherlands Nordic Singapore Spain Switzerland UK USA Other countries Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law. In general, the trademarks and designs referenced herein are trademarks, or registered trademarks, of Tecan Group Ltd., Männedorf, Switzerland. A complete list may be found at Product names and company names that are not contained in the list but are noted herein may be the trademarks of their respective owners. For technical details and detailed procedures of the specifications provided in this document please contact your Tecan representative V Tecan is in major countries a registered trademark of Tecan Group Ltd., Männedorf, Switzerland. 217 Tecan Trading AG, Switzerland, all rights reserved.