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Measurement of serum levels of natalizumab, an immunoglobulin G4 therapeutic monoclonal antibody
Authors:Rispens Theo  Leeuwen Astrid van  Vennegoor Anke  Killestein Joep  Aalberse Rob C  Wolbink Gerrit J  Aarden Lucien A
Affiliation:aCarlsberg Laboratory, 1799 Copenhagen V, Denmark;bDepartment of Biology, Illinois Wesleyan University, Bloomington, IL 61702, USA;cDepartment of Chemistry, Copenhagen University, 2100 Copenhagen Ø, Denmark;dDepartment of Chemistry and Biomolecular Sciences, Macquarie University, New South Wales 2109, Australia
Abstract:Chlorophyllide a is a metabolite late in the biosynthesis of chlorophylls and bacteriochlorophylls. Isolation procedures for chlorophyllide a from Rhodobacter capsulatus CB1200 and barley (Hordeum vulgare L.) are described and compared. R. capsulatus CB1200 is a double mutant in the bacteriochlorophyllide a biosynthetic pathway, and chlorophyllide a is excreted by the cells when grown in Tween 80-containing liquid medium. It was purified by liquid or solid phase extraction, yielding 7 mg of chlorophyllide a from 1 L of culture. In a second approach, intrinsic chlorophyllase activity was used to dephytylate chlorophyll in an acetonic preparation of leaves of wild-type or chlorophyll b-deficient barley. Purification was achieved by liquid phase extraction, yielding 14 μg of chlorophyllide a per gram of barley leaves. Chlorophyllide a was identified by thin layer chromatography, absorption spectroscopy, and mass spectrometry.
Keywords:Chlorophyll biosynthesis   Chlorophyllide a   Photodynamic therapy   Chlorophyllase   Rhodobacter capsulatus
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