(675d) Multi-Well Device for High-Throughput Screening of Any Flat Sheet Membrane Adsorber

The development of membrane chromatography processes for biomanufacturing requires extensive optimization of the process conditions (e.g. solution pH, solution ionic strength). While many previous studies have demonstrated the advantages of performing optimization studies using 96-well filter plates (e.g. AcroPrep Advance from Pall¹) either on manual systems or on robotic platforms, this approach is limited to the few membrane types that are sold in a filter plate format. There are many types of emerging membrane chromatography materials (e.g. 3D printed membranes², self-assembled block copolymer membranes³) that would greatly benefit from the ability to efficiently conduct a high-throughput screening (HTS) study of their separation performance.

Inspired by the recent trend of using rocker platforms for continuous perfusion process in organ-on-a-chip applications (e.g. IFlowPlate⁴), a HTS method for membrane chromatography involving laterally-fed, gravitational flow driven by a perfusion rocker was developed. The process is based on a custom multi-well plate with exterior dimensions and well-to-well spacing based on standard 96-well plate sizing, allowing for its integration with manual and automated liquid dispensers (e.g. multichannel pipettes). Each multi-well plate can be used to conduct up to 32 experiments in parallel, with each experiment requiring just a single small disc of membrane; additionally multiple devices can be stacked on the perfusion rocker to conduct more than 32 experiments at once. The continuous flow of solution back and forth across the membrane during each ‘stage’ of the chromatography process (i.e., bind, wash, elute) occurs due to passive liquid levelling in response to the interval rocking action. As a proof of principle, we first analyzed the binding capacity of commercial anion-exchange membranes (i.e. Mustang Q, Sartobind Q, Natrix Q) for typical biomolecules (e.g. BSA, DNA). Interestingly, we found a considerable disparity in the reported binding capacity values for these biomolecules in the current literature (e.g. BSA binding capacity for Sartobind Q has been reported as 18 and 29 mg/mL^5,6) which provided additional motivation for an effective HTS tool for membrane chromatography processes.

Overall, the multi-well device developed in this study will allow for an improved understanding of biomolecule binding onto existing and emerging chromatographic membrane materials, which in turn will result in the development of separation processes that yield superior performance.

References

1. Pall. AcroPrep^TM Advance 96-well Filter Plates. Accessed April 7, 2022. https://shop.pall.com/us/en/products/filter-plates/96-well-filter-plates
2. Dong Z, Cui H, Zhang H, et al. 3D printing of inherently nanoporous polymers via polymerization-induced phase separation. Nat. Commun. 2021;12(247):1-12. doi: 10.1038/s41467-020-20498-1
3. Hahn J, Clodt JI, Filiz V, et al. Protein separation performance of self-assembled block copolymer membranes . Biotechnol Bioeng. 2014;4(20):10252–10260. doi: 10.1039/c3ra47306f
4. Rajasekar S, Lin DSY, Abdul L, et al. IFlowPlate - A customized 384-well plate for the culture of perusable vascularized colon organoids. Advanced Materials, 2020. doi: 10.1002/adma.202002974
5. Sartorius. Operating Instructions: Sartobind 96-well plates. 2018. Accessed April 4, 2022. https://www.sartorius.com/download/984502/manual-sartobind-96-well-plate...
6. MilliporeSigma. Natrix Q Chromatography Membrane: Comparison of process-related impurity clearance for membrane adsorbers and resin-based chromatography technologies. 2020. Accessed April 4, 2022. https://www.sigmaaldrich.com/CA/en/technical-documents/technical-article...