Application Notes

As the demand for plasmid DNA (pDNA) based gene therapy and vaccines increases, large scale, cost effective, and reproducible pDNA production will be required. The key to success is a real time in-process control method that ensures a high percentage of supercoiled pDNA in the final product. CIMac™ pDNA Analytical Column allows the monitoring of degradation products (open circular and linear pDNA), the removal of impurities (RNA), and ensures that each production step is yielding the amount of supercoiled pDNA anticipated.

The demand for monoclonal antibodies is invariably increasing on an annual basis. To satisfy increasing demands, faster and cheaper ways of manufacturing are explored. A quest for alternative paths in manufacturing not only requires development of most economical manufacturing process, but also rapid method development and development of good analytics for monitoring of manufacturing. For a quickly developed process, the use of reliable and fast analytical techniques are crucial. Moreover, this analytical technique should than be preferably used also for in-process control during manufacturing stage.

Here we present fast and reliable method for processing and analyzing IgG, IgA ang IgM using CIM® QA Disk Monolithic Column, which thrive upon speed, repeatability and high capacity.

A Hemoglobin A1c reference standard was loaded on CIM® SO3 monolithic column and eluted in a mixed stepwise and linear gradient. HbA1a, HbA1b and HbA0 variants were separated and a complete determination of HbA1c (including equilibration) was obtained within 1.1 minute.

A mixture of IgG, HSA and IgM standard was loaded on CIM® EDA Disk and eluted in linear salt gradient at a flow rate of 4 mL/min (12 CV/min). A complete separation of IgM from IgG and HSA was obtained within 1.5 minute.

A supernatant from Phanerochaete chrysosporium cultivation was loaded on CIM® QA Disk, and elution was effected by a linear gradient at a flow rate of 3 mL/min (9 CV/min). Baseline separation of isoenzymes H2, H6/H7, H8 and H10 was achieved in less than 3 minutes.

Immunoaffinity columns were prepared by immobilization of Protein G on CIM® Epoxy Disk, CIM® Epoxy tube (1 mL) and an activated, particle based agarose support. A comparison of productivity was performed by loading centrifuged human plasma and resulted in superior productivity of CIM® monolithic supports.