Separation of specific immunoglobulin. 1. Desalination using a membrane system.
Separation of the ammonium sulfate precipitated protein fraction of mouse ascitic fluid, containing the specific immunoglobulin (pI 6.7-6.8; molecular weight 180000), from ammonium sulfate was investigated by means of non-traditional dialysis, based on the difference in diffusion rates of small and large molecules through porous membranes. The experiments were carried out in spiral membrane modules equipped with a Neosepta (AM-2 or ACS-SB) anion exchange membrane and a microfiltration membrane (Synpore or Sartorius). To enhance the driving force for penetration of ammonium sulfate and low-molecular-weight components from solution of ascitic protein fraction into water, a counterpressure was imposed on the side of microfiltration membrane. The flow rate, counterpressure and the pore sizes of microfiltration membranes had a significant effect on the separation process, as expected. The type of the anion exchange membrane had only a small effect. This process makes it possible to desalt the immunoglobulin fraction with high purity and yield in a few hours instead of 5 days.
Tishchenko
G
G
Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic.
Bleha
M
M
Skvor
J
J
Bures
L
L
D000645:Ammonium Sulfate; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001202:Ascitic Fluid / Q000276:immunology; D001709:Biotechnology; D003956:Dialysis / Q000295:instrumentation / Q000379:methods; D005069:Evaluation Studies as Topic; D005374:Filtration / Q000295:instrumentation / Q000379:methods*; D007136:Immunoglobulins / Q000302:isolation & purification*; D008567:Membranes, Artificial; D051379:Mice; D008851:Micropore Filters; D011233:Precipitin Tests; D012965:Sodium Chloride / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D008567:Membranes, Artificial; D012965:Sodium Chloride; D000645:Ammonium Sulfate