Publications
1996
A. Štrancar, P. Koselj, H. Schwinn, D. Josić
Analytical Chemistry, Vol. 68, No. 19, October 1, 1996
Production and downstream processing in biotechnology requires fast and accurate control of each step in the process. Improved techniques which can be validated are required in order to meet these demands. For these purposes, chromatographic units containing compact porous disks for fast separation of biopolymers were developed and investigated with regard to their performance and speed. The problems that have, in the past, arisen from the use of wide and flat separation units, such as membranes and disks, have chiefly been those of sample distribution and large void volumes before and behind the unit. Improvements in the construction of the cartridge have led to better performance of the compact porous disks and faster separation. Using these disks, three calibration standard proteins could be separated within less than 1 min by an anion-exchange, cation-exchange, and hydrophobic interaction mode. Such units can be used for in-process control in production and downstream processing of biopolymers, as was shown in experiments involving the purification of α1-antitrypsin and clotting factor IX and the immobilization of enzyme glucose oxidase on an epoxy-activated compact porous disk.
1995
B. G. Belenkii, V. G. Malt'sev
BioFeature, BioTechniques, 288, Vol. 18, No. 2 (1995)
In gradient chromatography for proteins migrating along the chromatographic column, the critical distance X0 has been shown to exist at which the separation of zones is at a maximum and band spreading is at a minimum. With steep gradients and small elution velocity, the column length may be reduced to the level of membrane thickness-about one millimeter. The peculiarities of this novel separation method for proteins, high-performance membrane chromatography (HPMC), are discussed and stepwise elution is shown to be especially effective. HPMC combines the advantages of membrane technology and high-performance liquid chromatography, and avoids their drawbacks.
1990
T. B. Tennikova, B. G. Belenkii, F. Švec
Journal of Liquid Chromatography, 13(1), 63-70 (1990)
Basing on the fact that only short layers of a chromatographic column contribute to the separation in the interaction chromatography, 1 mm thick membranes from macroporous methacrylate polymer provided with functional groups were synthetized and used for protein separation. The chromatograms show that the separation is fully comparable with that experienced on a filled column but the advantage of a membrane is up to two orders of magnitude lower pressure during the process and very high loading reaching up to 40 g/m2. This recommends the high performance membrane chromatography also for large scale preparative separations.