(340g) ChromaWeb™ platform, a linearly-scalable, high-resolution membrane chromatography cassette for purification of viral vectors.

Gene therapy treatments are expected to grow rapidly with at least 4 treatments already approved and many more in the pipeline. Therefore, there is a need for large-scale viral particle isolation without the need for ultracentrifugation or chemical methods. Chromatographic purification would be highly desirable, but the large size of viral vectors — 25~350 nm — requires adsorptive media having pores in the micron scale. This is the domain of adsorptive membranes and certain monoliths, but not of conventional chromatographic media whose pores are only in the 20~40 nm range.

Membranes have very fast kinetics by virtue of having very small diffusional distances because convective flow occurs through the adsorptive pores. Thus, the dynamic binding capacity is almost constant with a weak dependence on residence time, but with the inevitable tradeoff of lower binding capacity and high hydraulic resistance (HR) compared to conventional beads. The high HR demands very shallow beds of 5~15 mm, leading commercially available membrane chromatography devices to have relatively poor chromatographic resolution. Furthermore, none are linearly scalable. Therefore, there is a need for membrane chromatography devices that overcome these limitations.

The ChromaWeb modular chromatography platform is a stackable, planar, linearlyscalable cassette having a novel flow distribution network that delivers a very narrow residence time distribution (RTD) in spite of having a shallow bed and modules in parallel. As a result, ChromaWeb cassettes have HETPs on the order of 30 µm, at least 10 times smaller than the best commercially available devices, and peak asymmetry very close to 1 (vs. > 3). The modular and linearly-scalable structure of the ChromaWeb cassette enables results based on 10 mL lab scale cassettes to be directly translatable to clinical-scale (100~500 mL) and process-scale cassettes (1~10 L).

In this presentation we describe the structure of the ChromaWeb cassette, including its novel distribution network, and how it is capable of delivering linear-scalability and high resolution. Experimental results will be presented with a model protein on a HyperProductive ® bind-and-elute process at a 10·mL scale demonstrating very high productivity, high eluate concentration, and low buffer consumption.

The ChromaWeb platform is in its early development stage at SPF Technologies, and in search of commercial partners to bring it to market.