title: Reduction of strong lipase-polyclonal antibodies binding by limited proteolysis.
abstract: Initial attempts to purify Geotrichum candidum lipase (EC 3.1.1.3) using immunoaffinity chromatography have been hampered by the tight binding of the lipase to the immobilized rabbit anti-GCL IgG. Stringent elution conditions were unable to release more than a few percent of the antigen. To decrease the tight binding, the immunosorbent was treated with minute amounts of different proteases, of which elastase proved to be most effective. Using the elastase-treated immunosorbent both natural and recombinant GCL II were purified to homogeneity with 30% of lipase recovered from the immunoaffinity chromatography step.
authors.affiliation: National Research Council of Canada, Biotechnology Research Institute, Montreal, Quebec.
authors.forename: A
authors.initials: A
authors.lastname: Recktenwald
authors.forename: T
authors.initials: T
authors.lastname: Vernet
authors.forename: A C
authors.initials: AC
authors.lastname: Storer
authors.forename: E
authors.initials: E
authors.lastname: Ziomek
mesh_terms: D000908:Antibodies, Fungal / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D005848:Geotrichum / Q000201:enzymology*; D007164:Immunosorbents; D008049:Lipase / Q000276:immunology / Q000302:isolation & purification*; D010196:Pancreatic Elastase / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification; D013381:Subtilisins / Q000378:metabolism; D013820:Thermolysin / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D000908:Antibodies, Fungal; D000936:Antigen-Antibody Complex; D007164:Immunosorbents; D011994:Recombinant Proteins; D008049:Lipase; D010450:Endopeptidases; D013381:Subtilisins; D010196:Pancreatic Elastase; D014357:Trypsin; D013820:Thermolysin