Double-modified albumins as a tool for antibody purification.
A general approach for anti-hapten antibody purification utilizing double-modified albumins is presented. Purification is based on simultaneous modification of an albumin with a hapten (e.g. fluorescein) and desthiobiotin. Three distinct albumins (BSA, HSA and ovalbumin) were modified accordingly and evaluated for their ability to purify the anti-fluorescein mAb from a mixture of commercial preparation and an E. coli cell lysate. The recovered mAb was obtained at relatively high purity (88-95%), in a wide range of target concentrations (0.66-0.02 mg/ml) within a total purification time of approximately 20 min. Substantial increase in the contamination background did not affect purity.
Patchornik
Guy
G
Affisink Biotechnology Ltd., 11 Hamaccabee St. Kiryat-Ono 55572, Israel. guy@affisink.com
D000906:Antibodies / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D010320:Parvalbumins / Q000276:immunology* / Q000302:isolation & purification; D012709:Serum Albumin / Q000276:immunology* / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000276:immunology* / Q000302:isolation & purification
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D010320:Parvalbumins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine