Reduction of Strong Lipase-Polyclonal Antibodies Binding by Limited Proteolysis
✍ Scribed by A. Recktenwald; T. Vernet; A.C. Storer; E. Ziomek
- Book ID
- 102561826
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 310 KB
- Volume
- 226
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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.