title: Esterase activity of carbonic anhydrases serves as surrogate for selecting antibodies blocking hydratase activity.
abstract: Carbonic anhydrase 9 (CA9) and carbonic anhydrase 12 (CA12) were proposed as potential targets for cancer therapy more than 20 years ago. However, to date, there are only very few antibodies that have been described to specifically target CA9 and CA12 and also block the enzymatic activity of their targets. One of the early stage bottlenecks in identifying CA9- and CA12-inhibiting antibodies has been the lack of a high-throughput screening system that would allow for rapid assessment of inhibition of the targeted carbon dioxide hydratase activity of carbonic anhydrases. In this study, we show that measuring the esterase activity of carbonic anhydrase offers a robust and inexpensive screening method for identifying antibody candidates that block both hydratase and esterase activities of carbonic anhydrase's. To our knowledge, this is the first implementation of a facile surrogate-screening assay to identify potential therapeutic antibodies that block the clinically relevant hydratase activity of carbonic anhydrases.
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .
authors.forename: Narasimha Rao
authors.initials: NR
authors.lastname: Uda
authors.affiliation: b 4-Antibody AG (Wholly-Owned Subsidiary of Agenus Inc., USA) , Basel , Switzerland .
authors.forename: Volker
authors.initials: V
authors.lastname: Seibert
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .|c Department Internal Medicine , University Hospital Basel , Basel , Switzerland .
authors.forename: Frank
authors.initials: F
authors.lastname: Stenner-Liewen
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .
authors.forename: Philipp
authors.initials: P
authors.lastname: Müller
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .
authors.forename: Petra
authors.initials: P
authors.lastname: Herzig
authors.affiliation: d Department of Otorhinolaryngology , Klinikum der Universität München , Munich , Germany , and.|e Helmholtz Zentrum München - German Research Center for Environmental Health, Research Unit Gene Vectors , Munich , Germany.
authors.forename: Gabor
authors.initials: G
authors.lastname: Gondi
authors.affiliation: d Department of Otorhinolaryngology , Klinikum der Universität München , Munich , Germany , and.|e Helmholtz Zentrum München - German Research Center for Environmental Health, Research Unit Gene Vectors , Munich , Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Zeidler
authors.affiliation: b 4-Antibody AG (Wholly-Owned Subsidiary of Agenus Inc., USA) , Basel , Switzerland .
authors.forename: Marc
authors.initials: M
authors.lastname: van Dijk
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .|c Department Internal Medicine , University Hospital Basel , Basel , Switzerland .
authors.forename: Alfred
authors.initials: A
authors.lastname: Zippelius
authors.affiliation: a Department of Biomedicine , University of Basel , Basel , Switzerland .
authors.forename: Christoph
authors.initials: C
authors.lastname: Renner
mesh_terms: D000086:Acetazolamide / Q000737:chemistry / Q000494:pharmacology; D000154:Aconitate Hydratase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology*; D000951:Antigens, Neoplasm / Q000378:metabolism*; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004950:Esterases / Q000378:metabolism*; D006801:Humans; D015394:Molecular Structure; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D004791:Enzyme Inhibitors; D004950:Esterases; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases; C477378:carbonic anhydrase XII; D000154:Aconitate Hydratase; D000086:Acetazolamide
keywords: Cancer therapy; carbonic anhydrase 9 & 12; enzyme inhibition; high-throughput screening; renal cell cancer