Disposable, 96-well filter plates prefilled with chromatography resins are suited for high-throughput chromatography screening and optimization of resins and chromatographic conditions, including binding, wash, and elution conditions.
These 96-well plates offer an essential first step for characterization of optimal conditions used for large-scale purification of important biotherapeutics such as monoclonal antibodies (mAbs) and for the manufacture of vaccines. The plates can be used with a robotic system or operated manually with a multichannel pipette (using vacuum or centrifugation).
In this article, we present an overview of the workflow using PreDictor™ 96-well plates, together with a checklist of equipment required for running high-throughput process development (HTPD) experiments with these tools in a manual setup.
HTPD and parallel chromatography
Efficient development of the purification process in bioprocess workflows is a crucial component of overall biopharmaceutical development. HTPD shortens development time and at the same time increases the amount of information available during early bioprocess development.
HTPD workflow using PreDictor 96-well plates
The workflow using PreDictor plates is performed in a batch mode and comprises the same steps as any chromatography column experiment (see Fig 1):
- Equilibration of the resin with the desired buffer
- Sample application
- Wash
- Elution
We start the workflow with conditioning of the resin by repeated addition of equilibration/binding buffer. Sample in binding buffer is then added to the resin and incubation is started. All incubations, whether with buffer or sample, are performed with agitation on a shaker, while separation of phases is performed by centrifugation or vacuum filtration.
The sample solution is then removed by repeated washes with the chosen solution, followed by repeated elution with the chosen conditions.
Fig 1. Steps in batch purification using PreDictor 96-well plates.
Watch a demonstration of a manual operation using PreDictor plates.
Equipment checklist for getting started with PreDictor 96-well plates
Table 1. Equipment checklist
|
Equipment |
Detail |
|
PreDictor plate |
Select the appropriate PreDictor plate with the Purify App. A selection of our PreDictor plates can be found here. |
|
Collection plates |
Collection plate for 96-well plates (product code 28403943) (not for UV absorbance) |
|
UV-readable collection plates |
Example: Corning 96-well clear flat bottom UV-transparent microplate (3635) |
|
Buffer container (v-shaped) |
To be used when you want the same condition in all wells of the plate. Example: Eppendorf reagent reservoir, autoclavable reservoir for multichannel pipette, filling volume 50–60 mL (0030058607). |
|
Buffer/storage plates with 48 or 96 wells for buffer preparation |
Use buffer/storage plates with 48 or 96 (or even down to 8) wells to contain the respective buffers. Examples: |
|
Microplate foil |
Biacore™ microplate foil (product code 28975816) |
|
Multichannel pipette |
Use an 8- or 12-channel pipette for quick and easy pipetting of liquids into the PreDictor plates |
|
Vacuum manifold |
Multiwell plate manifold (product code 5017)
|
|
Microplate shaker |
Use a microplate shaker with 3 mm circular centripetal movement and regulation speed of 1100 rpm to fully suspend the sample/buffer in the resin during incubation. Example: IKA MTS 2/4 digital microtiter shaker. (3208000) |
|
Centrifuge |
Use a swing-out rotor with microplate carriers capable of handling a PreDictor plate on top of a collection plate |
Related resources
CY13346