ScreenExpert RoboColumn

Transcription

1 USD2992 (1) For High Throughput Chromatography Sorbent Screening and Optimization s are miniaturized chromatography columns pre-packed with Pall chromatography sorbents. They are designed for fully automated and parallel chromatographic separations with robotic liquid handling workstations, such as the Freedom EVO from Tecan. s are packed with Pall anion exchange, mixed-mode or any other Pall chromatography sorbents upon request. The column units are packed by Atoll GmbH and are identical to Atoll s MediaScout RoboColumns. Two formats are available, 2 µl and 6 µl, with a bed height of 1 and 3 mm respectively. The 2 µl column volume is recommended for sorbent and process conditions screening, while the 6 µl is used for purification process optimization or when a residence time above 4 minutes is required (e.g., capacity study on mixed-mode sorbents). The columns are supplied as rows of eight pre-packed units, with two removable silicon cover seals for proper storage. A 96-well array plate is available to arrange up to 96 ScreenExpert RoboColumn units. Refer to Table 1 for characteristics and available column formats.

2 Benefits Column format, and high quality packing enables the ability to mimic all the steps of a chromatography separation in dynamic mode. Data obtained are comparable to those obtained in larger columns Miniaturized format facilitates screening of chromatography sorbents, reduces sample consumption, and saves time Perfectly fitted to Design of Experiment (DoE) requirements, allowing a large amount of experimental work in a minimum of time 96-well format allows easy integration with automation systems for High Throughput Process Development (HTPD) Multiple parameters can be tested in parallel, e.g., residence time Reusable Flexible use: the user makes column selection when needed Characteristics Table 1 s Characteristics s Column volume 2 µl 6 µl Bed height 1 mm 3 mm Column inner diameter 5 mm Column material Polypropylene Chemical stability All commonly used aqueous buffers, ph 1 to 14 Avoid Halogenated organic solvents, hexane Storage solution 2% ethanol /1 M NaCl Recommended storage 2 to 8 C temperature Working temperature 4 to 3 C Maximum working pressure Up to 8. bar g (116 psi g) Flow rate 16 to 1, cm/hr (1) Laser label Placed on every individual column body, and containing: Product number Sorbent name Column volume in µl Applications s are suitable for a large variety of applications such as: Parallel screening of chromatography sorbents Parallel screening and optimization of chromatographic conditions in downstream process development (conductivity, ph) Scale down experimental work Determination of dynamic binding capacity Determination of the optimal residence time Optimization of the regeneration step Application Example: Development of the Capture Step of a Monoclonal Antibody (MAb) Expressed in a Crude CHO Feedstock MAb binding conditions were tested on 3 mixed-mode sorbents and dynamic binding capacity (DBC) was evaluated based on breakthrough (BT) curve (Figure 1). The was overloaded with MAb CHO feedstock at.52 mg/ml and flowthrough (FT) was collected as one-column-volume (CV) fractions (2 µl each) in 35 µl 96-well plate. The amount of MAb recovered in the FT fractions was evaluated using Bio-Layer Interferometry (BLI) with protein A sensor (Octet System, Pall ForteBio). It was plotted against the volume of feedstock loaded per unit. 1/1 of the MAb concentration (C/1) was.52 mg/ml (52 µg/ml in Figure 1). The volume equivalent at 1% BT (Veq1%BT) was the volume of MAb loaded when reaching C/1 in FT fractions (8.56 ml in Figure 1). 96-Well Array Plate Size x 86. x 14. mm Material Polyoxymethylene copolymer Number of wells 96 (1) The Liquid Transfer menu of the Te-Chrom Wizard enables set up of the flow rate for each individual dispensing and chromatography step ( Liquid Transfer, Te-Chrom Wizard Software Manual, 39676, en, V1.). 2

3 Figure 1 Breakthrough Curve on MEP HyperCel Sorbent in 2 µl Figure 3 DBC on MEP HyperCel Sorbent in and in PRC Column DBC 1% BT (mg MAb/mL Sorbent) PRC Column Breakthrough curve during the loading of a MAb CHO feedstock (.52 mg/ml) at a ~4-minute residence time. DBC1% BT (mg/ml sorbent) = Veq1%BT (ml)* x MAb concentration (mg/ml) Column volume (ml) *void volume was taken into account Using this breakthrough curve method, the DBC for MEP, HEA and PPA HyperCel sorbents was determined on 2 µl s (Figure 2). Figure 2 DBC on 3 Mixed-Mode HyperCel Sorbents in ScreenExpert RoboColumns for Different Binding Conditions This comparison showed that MAb binding was maximized on DBC1% BT (mg MAb/mL Sorbent) MEP HyperCel MEP HyperCel PBS, ph M NaCl, ph HEA HyperCel 1 M NaCl, ph 7.4 HEA HyperCel 1 M NaCl, ph 8.5 MEP HyperCel sorbent in PBS, ph 7.4. The selected condition was then transferred to 1 ml PRC column for confirmation of the DBC, approximately 21 mg/ml (Figure 3) PPA HyperCel PBS, ph 7.4 A 335-fold scale-up of the MAb capture step was finally carried out from 2 µl to LRC 25 lab-scale column (2.5 cm I.D) packed with 67 ml MEP HyperCel sorbent (Figure 4). Figure fold Scale-Up of the MAb Capture Step from ScreenExpert RoboColumn to LRC 25 Lab-Scale Column FT W E1 E2 E3 mau Abs. at 28 nm ph Conductivity ml Chromatogram of MAb Capture Step on LRC 25 lab-scale column (67 ml). Equilibration in PBS, ph 7.4. Loading of MAb feedstock = 21 mg MAb/mL MEP HyperCel sorbent. Residence time = 4 minutes; FT: Flowthrough; W: Wash; E1, E2 and E3: Elution at ph 5, 4 and 3 respectively. The analysis of fractions obtained on and LRC 25 lab-scale column were in the same magnitude. Comparable data in terms of recovery (1%) and purity (9%) were evidenced (Figure 5, Table 2). ph

4 Figure 5 Fraction Analysis From and LRC 25 Lab-Scale Column Using BLI Table 2 Summary of the MAb Capture Step on ScreenExpert RoboColumn or LRC 25 Lab-Scale Column MAb Loading Column Volume Loaded (mg MAb/ Recovery Purity Type (ml) (mg)* ml sorbent) (%)* (%)** ScreenExpert RoboColumn LRC 25 Lab Scale Column * Evaluated using BLI technology (Octet System, Pall ForteBio) ** Evaluated using LabChip GXII technology (PerkinElmer) Handling s are designed for robotic handling. When using the high-throughput process development approach for s, it is suggested to use Design of Experiments (DoE) criteria for the experimental set-up. This enables to screen many different chromatographic parameters such as resin type, ph, conductivity / ionic strength, etc., efficiently and simultaneously. For experimental set-up and the configuration of the 96-well array plate, the Te-Chrom Wizard is recommended. The Te-Chrom Wizard is a dialog-based graphic user interface offered by Tecan and based on the Freedom EVOware software. The Te-Chrom Wizard was developed for the use of s and provides the possibility to configure hardware, plate layout, and the chromatographic process as well as to set all process relevant chromatographic parameters, e.g., flow rate, volume without direct script writing. Interaction of the MAb with protein A sensor was measured using BLI. Signal is correlated to MAb concentration using a standard curve (data not shown). Residence time = 4 minutes. F: Sample load; FT: Flowthrough; W: Wash; E1, E2 and E3: Elution at ph 5, 4 and 3 respectively. 4

5 Ordering Information MEP HyperCel 2 µl, row of 8 HEA HyperCel 2 µl, row of 8 PPA HyperCel 2 µl, row of 8 HyperCel STAR AX 2 µl, row of 8 MEP HyperCel 6 µl, row of 8 HEA HyperCel 6 µl, row of 8 PPA HyperCel 6 µl, row of 8 HyperCel STAR AX 6 µl, row of 8 96-well RoboColumn array plate Part Number SR2MEP SR2HEA SR2PPA SR2STARAX SR6MEP SR6HEA SR6PPA SR6STARAX SR96WAP PRC Prepacked Columns PRC Column 5x5 MEP HyperCel, 1 ml PRC Column 8x1 MEP HyperCel, 5 ml PRC Column 5x5 HEA HyperCel, 1 ml PRC Column 8x1 HEA HyperCel, 5 ml PRC Column 5x5 PPA HyperCel, 1 ml PRC Column 8x1 PPA HyperCel, 5 ml PRC Column 5x5 HyperCel STAR AX, 1 ml PRC Column 8x1 HyperCel STAR AX, 5 ml Part Number PRC5X5MEPHCEL1 PRC8X1MEPHCEL1 PRC5X5HEAHCEL1 PRC8X1HEAHCEL1 PRC5X5PPAHCEL1 PRC8X1PPAHCEL1 PRCSTARAX1ML PRCSTARAX5ML Complementary Products AcroPrep ScreenExpert Plates MEP, HEA and PPA HyperCel Mixed-Mode Sorbents MEP HyperCel Mixed-Mode Sorbent HyperCel STAR AX Anion Exchange Sorbent Part Number 96WP3MIXM5 96WPMEP5 96WPSTARAX5 Multi-Well Plates (empty) AcroPrep Advance 96-Well Filter Plate, 1 ml, μm Supor membrane (5/pkg) AcroPrep Advance 96-Well Filter Plate for Ultrafiltration 35 μl, Omega 3K MWCO (1/pkg) μl, Omega 1K MWCO (1/pkg) μl, Omega 3K MWCO (1/pkg) μl, Omega 1K MWCO (1/pkg) 836 Corporate Headquarters Port Washington, NY, USA toll free (USA) phone biopharm@pall.com European Headquarters Fribourg, Switzerland +41 () phone LifeSciences.EU@pall.com Asia-Pacific Headquarters Singapore phone sgcustomerservice@pall.com Visit us on the Web at us at biopharm@pall.com International Offices Pall Corporation has offices and plants throughout the world in: Argentina, Australia, Austria, Belgium, Brazil, Canada, China, France, Germany, India, Indonesia, Ireland, Italy, Japan, Korea, Malaysia, New Zealand, Norway, Philippines, Poland, Russia, Singapore, South Africa, Spain, Sweden, Switzerland, Taiwan, Thailand, United Kingdom, and Vietnam. Distributors in all major industrial areas of the world. To locate the Pall office or distributor nearest you, visit The information provided in this literature was reviewed for accuracy at the time of publication. Product data may be subject to change without notice. For current information consult your local Pall distributor or contact Pall directly. 214, Pall Corporation. Pall,, Octet, Omega, Supor and HyperCel are trademarks of Pall Corporation. indicates a trademark registered in the USA and TM indicates a common law trademark. Filtration. Separation. Solution.is a service mark of Pall Corporation. MediaScout, RoboColumn and RoboColumns are trademarks of Atoll GmbH. LabChip is a trademark of PerkinElmer. Freedom EVO, Freedom EVOware, and Te-Chrom are trademarks of Tecan Group. 9/14, PDF, GN USD2992(1)