Delta Precision Ltd.
Expanded Bed Adsorption chromatography (EBA) allows clarification and direct product capture in one unit operation.
Upflow conditions achieve a fluidized bed, twice the volume of the settled bed. Crude, high cell density and highly-viscous feed streams such as cell culture, foods and plasma are injected upflow.
Cells, cell debris, host cell proteins (HCP's) and DNA leave the expanded bed, whilst the target molecule is bound to the media.
In order to facilitate a renaissance in EBA, columns built to current Good Manufacturing Practice (cGMP) standards had to be designed, be scalable, and be both feasible and economic to manufacture.
The user requirement specifications brought a number of design and manufacturing challenges to overcome:
The Biotechflow column overcame all the challenges set out in the user requirement specifications to produce a new column design for expanded bed adsorption chromatography
Current applications are biopharmaceutical production from crude, high cell density and highly-viscous feed streams including cell culture, foods and plasma.
These columns are now installed in cGMP facilities in Europe, Australia and the USA.
They are presently used in Phase II and Phase Ill production of biopharmaceuticals including MAb's.
Tests to evaluate the performance of the EBA 300 column for a monoclonal antibody (MAb) using Rhobust'' MAbDirect proteinA(4) were conducted at cGMP manufacturing facilities.
High cell-density harvest material was used under the following conditions:
Cell density: 60-90 million cells mL-1 (viability >8C%)
Antibody titre: 1.34 g L-1 (continuous cell culture, cumulative titre 5.8 g L-1 over 21 days;
Load ratio: 22 mg MAb mL-1 settled bed (both Protein A resins)
The results detailed in Tables 1, 2 and 3 show that in comparison to traditional clarification
and chromatography processes, the EBA 300 column and technology realized the following
Figure 1. Shows the chromatogram from the 300mm EBA column using Rhobust'MAbDirect proteinA5).