In this work pressure drop was investigated for an estimation of dynamic binding capacity (DBC) of proteins and plasmid DNA for monoliths with different pore sizes. It was demonstrated that methacrylate monolith surface area is reciprocally proportional to pore diameter and that pressure drop on monolith is reciprocally proportional to square pore size. Based on these facts mathematical formalism has been derived predicting that DBC is in linear correlation with the square root of pressure drop what was experimentally confirmed.
COBISS.SI-ID: 16493846
In this work methacrylate monoliths bearing different combination of hydrophobic and ion-exchange groups were prepared. Monolith bearing combination of grafted C4 methacrylate groups and DEAE groups was found to enable separation of open circular (OC) from supercoiled (SC) pDNA forms and RNA from pDNA. Pure pDNA dynamic binding capacity was 4.7 mg/ml under IEX conditions and 2.1 mg/ml under HIC conditions. Single step purification enabled removal of over 99% of RNA, host cell proteins (HCP) and genomic DNA (gDNA) demonstrating capacity to purify around 1.5 mg of pDNA/ml of monolith from cell lysate.
COBISS.SI-ID: 4199288
In this review brief introduction to the monoliths is given with the emphasize on the theory of separation of large molecules, particularly on a linear gradient elution and estimation of peak broadening.
COBISS.SI-ID: 4263032