Sofia Farkona1, Antoninus Soosaipillai2, Panagiota Filippou3, Dimitrios Korbakis3, Stefano Serra4, Felix Rückert5, Eleftherios P Diamandis6, Ivan M Blasutig7. 1. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada; Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada. 2. Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada. 3. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada; Department of Clinical Biochemistry, University Health Network, Toronto, Canada. 4. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada; Department of Pathology, Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. 5. Visceral, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus, Technical University of Dresden, Germany. 6. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada; Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada; Department of Clinical Biochemistry, University Health Network, Toronto, Canada; Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada. 7. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada; Department of Clinical Biochemistry, University Health Network, Toronto, Canada. Electronic address: iblasutig@cheo.on.ca.
Abstract
BACKGROUND: CUB and zona pellucida-like domain-containing protein 1 (CUZD1) was identified as a pancreas-specific protein and was proposed as a candidate biomarker for pancreatic related disorders. CUZD1 protein levels in tissues and biological fluids have not been extensively examined. The purpose of the present study was to generate specific antibodies targeting CUZD1 to assess CUZD1 expression within tissues and biological fluids. METHODS: Mouse monoclonal antibodies against CUZD1 were generated and used to perform immunohistochemical analyses and to develop a sensitive and specific enzyme-linked immunosorbent assay (ELISA). CUZD1 protein expression was assessed in various human tissue extracts and biological fluids and in gel filtration chromatography-derived fractions of pancreatic tissue extract, pancreatic juice and recombinant protein. RESULTS: Immunohistochemical staining of CUZD1 in pancreatic tissue showed that the protein is localized to the acinar cells and the lumen of the acini. Western blot analysis detected the protein in pancreatic tissue extract and pancreatic juice. The newly developed ELISA measured CUZD1 in high levels in pancreas and in much lower but detectable levels in several other tissues. In the biological fluids tested, CUZD1 expression was detected exclusively in pancreatic juice. The analysis of gel filtration chromatography-derived fractions of pancreatic tissue extract, pancreatic juice and recombinant CUZD1 suggested that the protein exists in high molecular weight protein complexes. CONCLUSION: This study describes the development of tools targeting CUZD1 protein, its tissue expression pattern and levels in several biological fluids. These new tools will facilitate future investigations aiming to delineate the role of CUZD1 in physiology and pathobiology.
BACKGROUND:CUB and zona pellucida-like domain-containing protein 1 (CUZD1) was identified as a pancreas-specific protein and was proposed as a candidate biomarker for pancreatic related disorders. CUZD1 protein levels in tissues and biological fluids have not been extensively examined. The purpose of the present study was to generate specific antibodies targeting CUZD1 to assess CUZD1 expression within tissues and biological fluids. METHODS:Mouse monoclonal antibodies against CUZD1 were generated and used to perform immunohistochemical analyses and to develop a sensitive and specific enzyme-linked immunosorbent assay (ELISA). CUZD1 protein expression was assessed in various human tissue extracts and biological fluids and in gel filtration chromatography-derived fractions of pancreatic tissue extract, pancreatic juice and recombinant protein. RESULTS: Immunohistochemical staining of CUZD1 in pancreatic tissue showed that the protein is localized to the acinar cells and the lumen of the acini. Western blot analysis detected the protein in pancreatic tissue extract and pancreatic juice. The newly developed ELISA measured CUZD1 in high levels in pancreas and in much lower but detectable levels in several other tissues. In the biological fluids tested, CUZD1 expression was detected exclusively in pancreatic juice. The analysis of gel filtration chromatography-derived fractions of pancreatic tissue extract, pancreatic juice and recombinant CUZD1 suggested that the protein exists in high molecular weight protein complexes. CONCLUSION: This study describes the development of tools targeting CUZD1 protein, its tissue expression pattern and levels in several biological fluids. These new tools will facilitate future investigations aiming to delineate the role of CUZD1 in physiology and pathobiology.
Authors: Agnieszka Magdalena Rygiel; Lara Sophie Unger; Franziska Lena Sörgel; Emmanuelle Masson; Ryotaro Matsumoto; Maren Ewers; Jian-Min Chen; Peter Bugert; Louis Buscail; Tomasz Gambin; Grzegorz Oracz; Maria Winiewska-Szajewska; Agnieszka Mianowska; Jarosław Poznanski; Joanna Kosińska; Piotr Stawinski; Rafał Płoski; Dorota Koziel; Stanisław Gluszek; Helmut Laumen; Fredrik Lindgren; J Matthias Löhr; Anna Orekhova; Vinciane Rebours; Jonas Rosendahl; Andrea Párniczky; Péter Hegyi; Akira Sasaki; Fumiya Kataoka; Yu Tanaka; Shin Hamada; Miklós Sahin-Tóth; Eszter Hegyi; Claude Férec; Atsushi Masamune; Heiko Witt Journal: Pancreatology Date: 2022-05-01 Impact factor: 3.977