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Immunological Identification of Fungal Species

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Human Fungal Pathogen Identification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1508))

Abstract

Immunodetection is described in this chapter as a technique for producing specific antibodies for antigen detection of the major human fungal pathogens. In the case of Candida spp., heat-killed cells are used to immunize mice over a couple of weeks and then splenocytes are isolated and further fused with myelomas to easily propagate the antibodies produced in the mice. The resulting antibodies follow a purification process where antibody levels and concentrations are determined. Fungal cells are also lysed to obtain whole cell extracts as a prior step for identification of antigens using immunoprecipitation. Finally, this method permits the production of specific antibodies against fungi and the identification of the respective antigens in an in vivo model.

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Acknowledgement

This work was supported by the European Union project FP7 “FUNGITECT” [602125] and FP7 Marie-Curie Initial Training Network “ImResFun” [MC-ITN-606786]. We also thank Andriy Petryshyn for technical support, Florian Zwolanek for helpful insights on immunization of mice, the Monoclonal Antibody Facility (MAF) at the Campus Vienna Biocenter (VBC) in Vienna, and Thomas Lion at the Children’s Cancer Research Institute (CCRI) for his suggestions and guidance.

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    Authors and Affiliations

    1. CCRI—Children’s Cancer Research Institute, Vienna, Austria

      Filomena Nogueira & Leonel Pereira

    2. Labdia—Labordiagnostik GmbH, Vienna, Austria

      Filomena Nogueira & Leonel Pereira

    3. Max F. Perutz Laboratories, MFPL—Department of Medical Biochemistry, Medical University of Vienna, Campus Vienna Biocenter, Dr. Bohr-Gasse 9, Vienna, 1030, Austria

      Filomena Nogueira, Fabian Istel, Leonel Pereira, Michael Tscherner & Karl Kuchler

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    1. Filomena Nogueira
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    Correspondence to Karl Kuchler .

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    1. Children’s Cancer Research Institute (CCRI), St.Anna Kinderkrebsforschung, Vienna, Austria

      Thomas Lion

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    Nogueira, F., Istel, F., Pereira, L., Tscherner, M., Kuchler, K. (2017). Immunological Identification of Fungal Species. In: Lion, T. (eds) Human Fungal Pathogen Identification. Methods in Molecular Biology, vol 1508. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6515-1_20

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