Application of compact porous tubes for preparative isolation of clotting factor VIII from human plasma.
Membranes as well as compact porous disks are successfully used for fast analytical separations of biopolymers. So far, technical difficulties have prevented the proper scaling-up of the processes and the use of membranes and compact disks for preparative separations in a large scale. In this paper, the use of a compact porous tube for fast preparative separations of proteins is shown as a possible solution to these problems. The units have yielded good results, in terms of performance and speed of separation. The application of compact porous tubes for the preparative isolation of clotting factor VIII from human plasma shows that this method can even be used for the separation of very sensitive biopolymers. As far as yield and purity of the isolated proteins are concerned, the method was comparable to preparative column chromatography. The period of time required for separation was five times shorter than with corresponding column chromatographic methods. Compact porous disks made of the same support material can also be used for in-process analysis in order to control the separation. The quick response, which is obtained from these units within 5 to 60 s, allows close monitoring of the purification process.
Strancar
A
A
BIA d.o.o., Teslova 30, Ljubljana, Slovenia.
Barut
M
M
Podgornik
A
A
Koselj
P
P
Schwinn
H
H
Raspor
P
P
Josić
D
D
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005164:Factor IX / Q000302:isolation & purification; D005169:Factor VIII / Q000302:isolation & purification*; D006801:Humans; D008567:Membranes, Artificial; D008768:Methylmethacrylates / Q000737:chemistry; D008854:Microscopy, Electron; D011137:Polystyrenes / Q000737:chemistry; D016062:Porosity; D012117:Resins, Synthetic / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
D008567:Membranes, Artificial; D008768:Methylmethacrylates; D011137:Polystyrenes; D012117:Resins, Synthetic; C020347:GMA-EDMA; D005169:Factor VIII; D005164:Factor IX; C003771:Amberlite XAD-2 resin