Single-step purification of immunotoxins containing a high ionic charge ribosome inactivating protein clavin by carboxymethyl high-performance membrane chromatography.
High-performance membrane chromatography (HPMC) and HPLC hydroxyapatite chromatography were compared for their efficiency in purifying immunotoxins (ITs) containing the ribosome-inactivating protein clavin, which is characterized by a high anionic charge and a low molecular mass. Both methods efficiently removed unreacted clavin from the conjugate crude mixture, but only the cation-exchange HPMC allowed efficient single-step separation of the unreacted monoclonal antibody (mAb) from ITs obtained by different coupling procedures.
Dosio
F
F
Dipartimento di Scienza e Tecnologia del Farmaco, University of Turin, Italy. dosio@pharm.unito.it
Arpicco
S
S
Canevari
S
S
Figini
M
M
Gastaldi
D
D
D000838:Anions; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D002412:Cations; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D005656:Fungal Proteins / Q000302:isolation & purification*; D000922:Immunotoxins / Q000302:isolation & purification*; D008970:Molecular Weight; D011500:Protein Synthesis Inhibitors / Q000302:isolation & purification*; D012260:Ribonucleases
D000838:Anions; D000911:Antibodies, Monoclonal; D002412:Cations; D005656:Fungal Proteins; D000922:Immunotoxins; D011500:Protein Synthesis Inhibitors; D012260:Ribonucleases; C100767:CLA protein, Aspergillus clavatus