An innovative monolithic column preparation for the isolation of 25 kilo base pairs DNA

• A conical monolith is tailored for the isolation of 25 kbp DNA.
• A conical monolith with zero ‘wall channel’ is produced.
• Elimination of ‘wall channel’ maximises DNA recovery.
• Elimination of ‘wall channel’ is based on height adjustment technique.
• Wall channel size can be predicted based on column backpressure.

The use of large DNAs in preparing multivalent vaccines that will eventually give protective immunity against multiple pathogenic microbes is becoming a major debate nowadays. One of the important issues in ensuring the successful implementation of the new vaccine technology is the development of a chromatographic technique that can handle larger DNAs. This paper reports the development of a novel conical monolithic column format with pore and surface characteristics engineered for the isolation of 25 kbp DNA in a single step fashion. An effective method of eliminating wall channelling, a defect of most conventional monolithic chromatography systems which has caused significant loss of product, was applied to maximise DNA recovery. This method was based on a systematic reduction of wall channel size based on a predetermined correlation between column's back pressure and wall channel size of a particular monolith pore size.