Showing papers by "BIA Separations (Slovenia) published in 2004"
TL;DR: The suitability of methacrylate based anion exchange monolithic supports for the separation and purification of plasmid and genomic DNA has been explored and under optimal conditions recoveries exceeding 80% were obtained even for genomic DNA.
Abstract: The suitability of methacrylate based anion exchange monolithic supports for the separation and purification of plasmid and genomic DNA has been explored. The effect of the size of the channels, ionic strength of the solution, and ligand density on the dynamic binding capacity has been investigated. The dynamic binding capacity was found to be flow independent, at least up to a linear velocity of 700 cm h(-1), and exceeded 9 mg mL(-1) for all types of DNA. The recovery depends on the pH value of the mobile phase and its ionic strength as well as on the density of the active groups. Under optimal conditions recoveries exceeding 80% were obtained even for genomic DNA. Finally, the suitability of this approach is demonstrated by purification of a real-life sample.
93 citations
TL;DR: It will be demonstrated that plasmid DNA can be purified from bacterial cells using alkaline lysis followed by chromatography on a very short weak anion-exchange chromatographic columns-disks-with good purity and quality within a short time.
76 citations
TL;DR: In this work, methacrylate-based monolithic columns with the bed volume up to 8000 ml are characterized and are found to be incompressible under these operating conditions and resistant to strong alkaline conditions.
75 citations
TL;DR: The kinetics of immobilization was studied in detail for trypsin under dynamic conditions and revealed that the enzyme immobilized via imidazole carbamate groups already reached its highest activity in 5 min, in contrast, a much longer time was required for immobilization via epoxy groups.
Abstract: Monolithic Convective Interaction Media (CIM) have been activated with epoxide and imidazole carbamate functionalities and used as supports for covalent immobilization of protein A, deoxyribonuclease I, and trypsin. The efficiency of immobilization for these proteins was determined from the amount of bound IgG, degradation of DNA, and hydrolysis of Nalpha-benzoyl-L-arginine ethyl ester, respectively. The respective biological activities of trypsin and the binding capacity of protein A immobilized via imidazole carbamate groups were 11.45 and 2.25 times higher than those obtained for epoxide matrix while they were practically equal for deoxyribonuclease I. The kinetics of immobilization was studied in detail for trypsin under dynamic conditions and revealed that the enzyme immobilized via imidazole carbamate groups already reached its highest activity in 5 min. In contrast, a much longer time was required for immobilization via epoxy groups.
66 citations
TL;DR: It has been demonstrated that ToMV, which had been diluted considerably below the sensitivity of ELISA, was concentrated by several orders of magnitude in the one-step procedure, and may provide a much faster and more efficient way to concentrate highly diluted plant viruses.
60 citations
TL;DR: P pH gradient showed better MnP isoform separation comparing to the salt gradient, while application of combined pH-salt gradient resulted in further improvement.
18 citations