CCTC (Continuous Countercurrent Tangential Chromatography) Overview

CCTC purification can be described as the typical process steps found in column chromatography happening simultaneously, with the notable exception of sanitization due to it’s closed-loop nature. In the pipe diagram above an initial capture step is modeled. Each step represents a new buffer condition and the stage inside those steps is comparable to an individual column volume of buffer loaded on to a column. The washout of buffer from one step to another is dictated by the number of stages in the step and gamma the ratio of flow between the retentate and permeate of the step. To ensure complete washout of each step 3 – 5 stages are used in each non-binding step.

Steady-State Elution Facilitates Continuous Neutralization

The continuous nature of the CCTC produces a consistent product concentration and buffer molarity allowing for inline neutralizations as quickly as 2 minutes after low pH contact.

CCTC Flowpath

Each stage of the CCTC flowpath consists of a membrane connected in series by retentate and permeate tubing. We use a variety of membranes from Repligen with pore sizes ranging 1 – 4 um and in areas that scale as small as 10 cm2 to as large as 10.4 m2, which provide both excellent sizing for process development and manufacturing purifications.

Pictured below - the Repligen Hollow Fiber Filters we use in our systems.

ChromaTan has been hard at work developing 3D printed devices that use the fluid dynamic effect Dean’s Vortex Separation (DVS) to supplement membranes used on the CCTC. DVS devices can be cleaned and reused more easily than membranes making them a great tool for process development purification.

Dramatic Increase in Resin Productivity

With short cycle times and a dilute resin slurry the grams of product produced per volume of per hour is typically >10x the productivity of standard column chromatography.

CCTC Software Control

For process development operations we have developed an HMI with built in automation and the ability to modify all inputs manually giving users the tools to tailor their operation precisely to their needs. Calibrate, operate, and monitor all on one easy to use screen. For a view of the data trends for pressure, pH, conductivity, UV, and flowrate the data historiographer provides several customizable plots to make data comparison quick and straightforward.

Efficiently Cycle Resin

Cycle times between 20-50 minutes mean that the CCTC can be scaled to reach maximum cycling of the resin once the campaign is completed.

CCTC Resin Selection

A comparison of offerings from Purolite Resins

The CCTC is a flexible platform that can use many off-the-shelf resins such a MabSelect® and POROS® resin beads to provide direct comparison of performance to standard column chromatography. We have also partnered with Purolite® to develop custom resin for CCTC applications. For standard columns small beads with low cross-linking cannot be used due to pressurization problems that would arise from the resin over-packing. With the CCTC, the resin is suspended allowing the structure of the resin bead to be small, increasing the proportional surface area of each bead, and reducing the need for dense cross-linking, which increases the perfusability of the resin beads. These advantages can lead to shorter binding times and increased binding capacity.

More About Purolite

Purolite is the world’s premier resin-based separation, purification and extraction technology manufacturer and innovation leader, with manufacturing facilities, advanced research laboratories and over 1100 people employed world-wide.

Purolite brings innovative thinking and a distinguished history of resin technology expertise to the global Life Sciences marketplace. We provide APIs, enzyme carriers, immobilized enzymes, and chromatographic resins of the highest quality, to support research and development and production-scale applications in global pharmaceutical, food and beverage, bioprocessing, cosmetics and fine chemical markets.

Purolite's Jetting technology has been pivotal to many of our technological breakthroughs and developments at ChromaTan.

Jetting technology is a, patented method that produces uniform-size agarose beads. This offers unique performance characteristics over traditional resins for the purification of recombinant proteins and monoclonal antibodies.

A non-solvent process, Jetting has the added benefit of being more environmentally friendly when compared to traditional manufacturing methods.

As the only supplier of process-scale volumes of these uniform agarose beads, Purolite helps us achieve:
- Higher flow velocities
- Increased resin lifetime
- Higher productivity
- Reduced costs

Interested in learning more about our protein purification solutions?

Call or email us to learn more about system performance, applications, integration, and case studies.