2D/3D Chromatography — Two or Three Steps. One Run

In preparative 2D chromatography, the outlet of the first column is connected directly to a second purification column. On the Contichrom® CUBE, ChromIQ® transfers the selected eluate when the Dimension 1 UV signal meets the Product Transfer Window defined in ChromaCraft™. The system can apply the specified in-line dilution during transfer before Dimension 2 performs its own binding, washing and elution steps within the same automated method.

No. Comprehensive analytical 2D-LC normally transfers or repeatedly samples the first-dimension effluent into a rapid second-dimension separation to increase analytical peak capacity.

The Contichrom® process is an integrated preparative workflow. It transfers a defined product window from the first purification column to a second preparative column to continue product purification without an intermediate hold or manual reload.

All 2D configurations require the Column Bypass Valve Kit, which allows the system to flush or sanitise between dimensions without dismounting columns. Gradient-to-gradient 2D methods additionally require the Buffer Selection Valve Kit for Gradient Pump 1A & 1B, which enables independent eluent selection for each dimension’s gradient pump. 3D methods require all of the above plus the Column Selection Valve Kit, which enables the third column position. All accessories are fully integrated into ChromaCraft™ and ChromIQ® as standard — no additional configuration required.

No. The columns and resins used in your existing batch method are the starting point. The same Protein A column you use for batch capture becomes Dimension 1 in a 2D process. Your existing IEX or RP polishing column becomes Dimension 2. No new column procurement is required to configure a 2D method.

The Product Transfer Window is a ChromaCraft™ step configuration that defines the criteria for transferring the eluate from Dimension 1 to Dimension 2. You set a UV absorbance threshold — when the Dimension 1 UV trace crosses that threshold during elution, ChromIQ® diverts the eluate to Dimension 2. In-line dilution is applied simultaneously at the defined ratio. The transfer is fully automated — no manual action is required at the switchover point.

Often yes — particularly for isocratic-to-gradient transfers where the Dimension 1 elution buffer (e.g., low-pH Protein A elution buffer) is not compatible with optimal binding conditions on Dimension 2 (e.g., IEX). The CUBE’s pump network handles this in-line, applying the dilution you define in the Product Transfer Window. You set the ratio; the system does the rest.

Yes. The accessory valve kits expand capability without removing any existing functionality. The CUBE with a Column Bypass Valve Kit runs all standard 1D batch modes (isocratic, gradient, alternating batch), all 2D integrated batch configurations, and all continuous processes (CaptureSMB, MCSGP, N-Rich) on the same hardware. Process mode is selected in ChromaCraft™ or the relevant wizard — the hardware configuration handles the rest.

They are complementary but operate differently, and importantly, a single Contichrom® CUBE can run either a twin-column continuous process (CaptureSMB, MCSGP, or N-Rich) or a 2D/3D integrated batch process — not both simultaneously. Full downstream integration combining, for example, a CaptureSMB capture step with a 2D polishing-polishing train, or a CaptureSMB capture step with MCSGP polishing, is achieved using two CUBE systems operating in series — with viral inactivation or another required hold step between them. This architecture delivers the benefits of both continuous capture and multi-step automated polishing across a complete downstream train.

Yes for 2D, no for 3D. 2D methods developed on the Contichrom® CUBE transfer directly to the Contichrom® PILOT 300X (up to 300 mL/min) and TWIN LPLC and HPLC production systems. The same ChromIQ® method logic, Product Transfer Window parameters, and in-line dilution ratios apply at larger column scale — no method redevelopment required. 3D methods are not natively supported on the larger scale systems, though customized solutions are possible – please enquire.