Open-source milligram-scale, four channel, automated protein purification system.

Publication date: Feb 23, 2024

Liquid chromatography purification of multiple recombinant proteins, in parallel, could catalyze research and discovery if the processes are fast and approach the robustness of traditional, “one-protein-at-a-time” purification. Here, we report an automated, four channel chromatography platform that we have designed and validated for parallelized protein purification at milligram scales. The device can purify up to four proteins (each with its own single column), has inputs for up to eight buffers or solvents that can be directed to any of the four columns via a network of software-driven valves, and includes an automated fraction collector with ten positions for 1. 5 or 5. 0 mL collection tubes and four positions for 50 mL collection tubes for each column output. The control software can be accessed either via Python scripting, giving users full access to all steps of the purification process, or via a simple-to-navigate touch screen graphical user interface that does not require knowledge of the command line or any programming language. Using our instrument, we report milligram-scale, parallelized, single-column purification of a panel of mammalian cell expressed coronavirus (SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-229E) trimeric Spike and monomeric Receptor Binding Domain (RBD) antigens, and monoclonal antibodies targeting SARS-CoV-2 Spike (S) and Influenza Hemagglutinin (HA). We include a detailed hardware build guide, and have made the controlling software open source, to allow others to build and customize their own protein purifier systems.

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Concepts Keywords
Chromatography Automated
Influenza Channel
Oc43 Chromatography
Python Column
Simple Cov
Milligram
Open
Parallelized
Positions
Purification
Sars
Scale
Single
Source
Tubes

Semantics

Type Source Name
disease VO time
disease VO report
disease VO device
drug DRUGBANK Pentaerythritol tetranitrate
disease IDO process
disease IDO cell
disease MESH Influenza
drug DRUGBANK Coenzyme M
disease IDO production
pathway REACTOME Reproduction
drug DRUGBANK Alpha-1-proteinase inhibitor
disease IDO replication
drug DRUGBANK Aluminium
drug DRUGBANK Ethanol
drug DRUGBANK Isopropyl Alcohol
drug DRUGBANK Silicon
disease VO protocol
drug DRUGBANK Tenocyclidine
disease VO volume
drug DRUGBANK Phosphate ion
drug DRUGBANK Isoxaflutole
disease VO company
drug DRUGBANK Ilex paraguariensis leaf
drug DRUGBANK Etodolac
disease VO syringe
drug DRUGBANK Puromycin
disease VO viability
drug DRUGBANK Imidazole
drug DRUGBANK Citric Acid
drug DRUGBANK Tromethamine
drug DRUGBANK Glycerin
disease VO frequency
drug DRUGBANK Sodium lauryl sulfate
drug DRUGBANK Ademetionine
drug DRUGBANK Cysteamine

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