Preparation of Anti-protein and Anti-mannan Antisera against Fungal Cell Wall by Affinity Chromatography |
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| Affiliation: | 1. HIV Pathogenesis Section, Laboratory of Immunoregulation, NIAID/NIH, Rockville, MD, USA;2. Amsterdam University Medical Centers, University of Amsterdam, Infectious Diseases, Amsterdam, the Netherlands;3. Amsterdam Institute for Infection and Immunity, Amsterdam, the Netherlands;4. Amsterdam Institute for Global Health and Development, Amsterdam, the Netherlands;5. Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, USA;6. Karsh Division of Gastroenterology & Hepatology, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, CA, USA;7. F. Widjaja Inflammatory Bowel Disease Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA;8. HIV Monitoring Foundation, Amsterdam, the Netherlands;9. Public Health Service of Amsterdam, Infectious Diseases, Amsterdam, the Netherlands;10. Leiden University Medical Center, Center for Proteomics & Metabolomics, Leiden, the Netherlands;11. Amsterdam UMC, University of Amsterdam, Experimental Immunology, Amsterdam, the Netherlands;12. Amsterdam University Medical Centers, University of Amsterdam, Department of Global Health, Meibergdreef 9, Amsterdam, the Netherlands;1. Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia;2. Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands;3. Genos Glycoscience Research Laboratory, BIOCentar, Zagreb, Croatia;4. University of Exeter, Exeter, UK;5. Division of Gastroenterology, S. Camillo-Forlanini Hospital, Circonvallazione Gianicolense, Rome, Italy;6. Division of Gastroenterology, Casa Sollievo della Sofferenza Hospital, Istituto di Ricovero e Cura a Carattere Scientifico, San Giovanni Rotondo, Italy;7. Division of Gastroenterology, University Hospital, Padua, Italy;8. Humanitas University, Inflammatory Bowel Disease Center, Department of Gastroenterology, Humanitas Clinical and Research Hospital, Milan, Italy;9. F. Widjaja Foundation, Inflammatory Bowel and Immunobiology Research Institute, Cedars-Sinai Medical Center, Los Angeles, California;10. Division of Gastroenterology, University Hospital Azienda Ospedaliero-Universitaria Careggi, Florence, Italy;11. Valiant Clinic, Dubai, United Arab Emirates;1. College of Food Science and Technology, Huazhong Agricultural University, National Research and Development Center for Egg Processing, Wuhan, Hubei 430070, PR China;2. Department of Animal Science, Iowa State University, Ames, IA 50011, USA;1. Center for Molecular Prediction of Inflammatory Bowel Disease (PREDICT), Department of Clinical Medicine, Aalborg University, Copenhagen, Denmark;2. Section of Gastroenterology, Medical Department, Zealand University Hospital, Køge, Denmark;3. Department of Epidemiology Research, Statens Serum Institut, Copenhagen, Denmark;4. Department of Gastroenterology & Hepatology, Aalborg University Hospital, Aalborg, Denmark;5. INSERM, Institut Pierre Louis d’Epidémiologie et de Santé Publique, Sorbonne Universite, Paris, France;3. From the Department of Rheumatology, Erasmus University Medical Center, 3000 CA Rotterdam, The Netherlands;;4. Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands;;5. Department of Rheumatology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands;;6. Division of BioAnalytical Chemistry, VU University Amsterdam, 1081 HV Amsterdam, The Netherlands; |
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| Abstract: | Iranzo, M., Marcilla, A., Elorza, M. V., Mormeneo, S., and Sentandreu, R. 1994. Preparation of anti-protein and anti-mannan antisera against fungal cell wall by affinity chromatography. Experimental Mycology 18, 159-167. A novel and easy chromatographic method has been developed for the isolation of anti-protein and anti-mannan antisera from a population of polyclonal antibodies obtained against Candida albicans and Yarrowia lipolytica cell wall mannoproteins. The technique is based on the immobilization of mannan (to be used as immunoadsorbent) by Affi-Prep Hz resin after the oxidation of neighboring hydroxyl groups of the polysaccharide with sodium periodate. For Y. lipolytica polyclonal antiserum, a single chromatographic step using the homologous mannan was sufficient to obtain an antiprotein antibody preparation free of antimannan antibodies. For C. albicans, three chromatographic processes using homologous and heterologous mannan were needed to obtain a satisfactory antiprotein antiserum. The potential application of the anti-protein antiserum obtained has been demonstrated by indirect immunofluorescence assays of whole cells and electrophoretic analysis of wall proteins in C. albicans and Saccharomyces cerevisiae . |
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