FPLC purification of mouse monoclonal antibodies from ascitic fluid using blue DEAE and thiophilic sorbents.

Two fast methods for the purification of mouse monoclonal antibodies from ascites, fluids using Blue-DEAE and 'thiophilic' adsorbent (T gel) in the FPLC system are described. Blue-DEAE chromatography is useful only for IgG1, IgG2a and IgG2b monoclonal antibody purification. T gel is a satisfactory adsorbent for IgG2b purification. Other IgG subclasses and IgM can also be obtained by simple one-step procedures, but the preparations contain small amount of high molecular weight contaminants.

Preparation of ferric adsorbent paper and its interaction with phosphate-containing biomolecules.

A procedure for the synthesis of a selective adsorbent for phosphate-containing biomolecules is described. The sorbent is based on Whatman chromatography paper, which is activated with epichlorohydrine, followed by the coupling of iminodiacetic acid to the active surface of the sorbent. The immobilized complex-forming chelating groups are saturated with ferric ions. The synthesized adsorbent is a counterpart to Chelating Sepharose and makes it possible to extend the use of immobilized ferric chelating groups for analytical purposes. It displays a high affinity towards compounds containing free terminal phosphate groups (phosphopeptides, nucleotides). The results of the binding experiments are compared to the corresponding data obtained with Chelating Sepharose gels.