DataApex has produced a white paper providing application examples of its Clarity software for analytical and small-scale preparative separation using multivendor analytical and preparative HPLC components.
Introduction
Besides its use in analytical GC, HPLC and CE, the Clarity chromatography software is suited for control of preparative chromatography systems. Using the FCGP (General Purpose Fraction Collector) control module, many different fraction collectors can be controlled by the simple Next and Collect/Waste commands generated by the module and transferred to the devices either through the digital Outputs/Inputs or by a suitable communication line (RS232, LAN, GSIOC).
Two examples of automated systems used for isolation and purification of antibodies, based on common instrumentation and controlled by Clarity chromatography software are presented.
Automated immunoaffinity chromatography
This instrument setup is designed to isolate specific antibodies from clarified serum. The serum is obtained from an animal immunized with the antigen against which antibodies need to be generated. The media in the column has the same or a similar antigen covalently bound to it, in order to capture antibodies which are specific to that antigen.
Preparative size-exclusion chromatography with stacked injections
This instrument setup is designed to facilitate removal of aggregates and other high molecular weight species from antibodies. Generally, the harshness of the elution buffer used in the affinity step leads to some degree of aggregation in the isolated antibodies.
Conclusion
The use of Clarity is demonstrated in the purification of antibodies using two specific setups: the automated immunoaffinity chromatography and preparative size-exclusion chromatography with stacked injections. Similar setups can be used with a large range of fraction collectors and multi-position valves. The description can also help with setting up systems for other preparative applications.