For High Throughput Chromatography Sorbent Screening and OptimizationScreenExpert RoboColumns are miniaturized chromatography columns pre-packed with 颇尔 chromatography sorbents.They are designed for fully automated and parallel chromatographic separations with robotic liquid handling workstations, such as the Freedom EVO♦ from Tecan.
ScreenExpert RoboColumns are packed with 颇尔 anion exchange, mixed-mode or any other 颇尔 chromatography sorbents upon request.The column units are packed by Atoll GmbH and are identical to Atoll’s MediaScout♦ RoboColumns.
Two formats are available, 200 μL and 600 μL, with a bed height of 10 and 30 mm respectively.The 200 μL column volume is recommended for sorbent and process conditions screening, while the 600 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.
- 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
- Flexible use:the user makes column selection when needed
HandlingScreenExpert RoboColumns are designed for robotic handling.
When using the high-throughput process development approach for ScreenExpert RoboColumns, 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 ScreenExpert RoboColumns 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.
♦ MediaScout and RoboColumn are trademarks of Atoll GmbH.LabChip is a trademark of PerkinElmer.Freedom EVO, Freedom EVOware, and Te-Chrom are trademarks of Tecan Group.
- 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 ScreenExpert RoboColumn was overloaded with MAb CHO feedstock at 0.52 mg/mL and flowthrough (FT) was collected as one-column-volume (CV) fractions (200 μL each) in 350 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, 颇尔 ForteBio).It was plotted against the volume of feedstock loaded per ScreenExpert RoboColumn unit.1/10 of the MAb concentration (C/10) was 0.052 mg/mL (52 μg/mL in Figure 1).The volume equivalent at 10% BT (Veq10%BT) was the volume of MAb loaded when reaching C/10 in FT fractions (8.56 mL in Figure 1).
Breakthrough Curve on MEP HyperCel Sorbent in 200 μL ScreenExpert RoboColumn
Breakthrough curve during the loading of a MAb CHO feedstock (0.52 mg/mL) at a ~4-minute residence time.
DBC10% BT (mg/mL sorbent) =
|Veq10%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 200 μL ScreenExpert RoboColumns (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 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).
DBC on MEP HyperCel Sorbent in ScreenExpert RoboColumn and in PRC Column
A 335-fold scale-up of the MAb capture step was finally carried out from 200 μL ScreenExpert RoboColumn to LRC 25 lab-scale column (2.5 cm I.D) packed with 67 mL MEP HyperCel sorbent (Figure 4).
335-fold Scale-Up of the MAb Capture Step from ScreenExpert RoboColumn to LRC 25 Lab-Scale Column
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 ScreenExpert RoboColumn and LRC 25 lab-scale column were in the same magnitude.Comparable data in terms of recovery (100%) and purity (90%) were evidenced (Figure 5, Table 2).
Fraction Analysis From ScreenExpert RoboColumn and LRC 25 Lab-Scale Column Using BLI
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.F0:Sample load; FT:Flowthrough; W:Wash; E1, E2 and E3:Elution at pH 5, 4 and 3 respectively.
Table 2Summary of the MAb Capture Step on ScreenExpert RoboColumn or LRC 25 Lab-Scale Column
LRC 25 Lab
* Evaluated using BLI technology (Octet System, 颇尔 ForteBio)
** Evaluated using LabChip♦ GXII technology (PerkinElmer)
|Column volume||200 μL||600 μL|
|Bed height||10 mm||30 mm|
|Column inner diameter||5 mm|
|Chemical stability||All commonly used aqueous buffers, pH 1 to 14|
|Avoid||Halogenated organic solvents, hexane|
|Storage solution||20% ethanol /1 M NaCl|
|Recommended storage temperature||2 to 8 °C|
|Working temperature||4 to 30 °C|
|Maximum working pressure||Up to 8.0 bar g (116 psi g)|
|流速||16 to 1,000 cm/hr 1|
Placed on every individual column body, and containing:
|96-Well Array Plate|
|尺寸||128.3 x 86.0 x 14.0 mm|
1 The Liquid Transfer menu of the Te-Chrom♦ Wizard enables set up of the flow rate for each individual dispensing and chromatography step (220.127.116.11 Liquid Transfer, Te-Chrom Wizard Software Manual, 396076, en, V1.0).
- 使用说明：ScreenExpert RoboColumns
- 应用说明：Use of ScreenExpert RoboColumns for High Throughput Study of Loading Conditions on HyperCel™ STAR AX and MEP HyperCel Sorbents for MAb Purification in Flow-Through Mode
- 应用说明：High Throughput Regeneration Study on MEP HyperCel™ Mixed-Mode Sorbent on ScreenExpert RoboColumns
|ScreenExpert RoboColumn MEP HyperCel 200 μL, row of 8||SR2MEP|
|ScreenExpert RoboColumn HEA HyperCel 200 μL, row of 8||SR2HEA|
|ScreenExpert RoboColumn PPA HyperCel 200 μL, row of 8||SR2PPA|
|ScreenExpert RoboColumn HyperCel STAR AX 200 μL, row of 8||SR2STARAX|
|ScreenExpert RoboColumn MEP HyperCel 600 μL, row of 8||SR6MEP|
|ScreenExpert RoboColumn HEA HyperCel 600 μL, row of 8||SR6HEA|
|ScreenExpert RoboColumn PPA HyperCel 600 μL, row of 8||SR6PPA|
|ScreenExpert RoboColumn HyperCel STAR AX 600 μL, row of 8||SR6STARAX|
|96-well RoboColumn array plate||SR96WAP|
AcroPrep ScreenExpert Plates
|MEP, HEA and PPA HyperCel Mixed-Mode Sorbents||96WP3MIXM50|
|MEP HyperCel Mixed-Mode Sorbent||96WPMEP50|
|HyperCel STAR AX Anion Exchange Sorbent||96WPSTARAX50|
Multi-Well Plates (empty)
|AcroPrep Advance 96-Well Filter Plate, 1 mL, 0.45 μm Supor® membrane (5/pkg)||8129|
AcroPrep Advance 96-Well Filter Plate for Ultrafiltration
|350 μL, Omega™ 3K MWCO (10/pkg)||8033|
|350 μL, Omega 10K MWCO (10/pkg)||8034|
|350 μL, Omega 30K MWCO (10/pkg)||8035|
|350 μL, Omega 100K MWCO (10/pkg)||8036|
PRC Prepacked Columns
|PRC Column 5x50 MEP HyperCel, 1 mL||PRC05X050MEPHCEL01|
|PRC Column 8x100 MEP HyperCel, 5 mL||PRC08X100MEPHCEL01|
|PRC Column 5x50 HEA HyperCel, 1 mL||PRC05X050HEAHCEL01|
|PRC Column 8x100 HEA HyperCel, 5 mL||PRC08X100HEAHCEL01|
|PRC Column 5x50 PPA HyperCel, 1 mL||PRC05X050PPAHCEL01|
|PRC Column 8x100 PPA HyperCel, 5 mL||PRC08X100PPAHCEL01|
|PRC Column 5x50 HyperCel STAR AX, 1 mL||PRCSTARAX1ML|
|PRC Column 8x100 HyperCel STAR AX, 5 mL||PRCSTARAX5ML|