Use of phage antibodies to distinguish closely related species of protozoan parasites

Article type: Research Article

Authors: Timothy Paget, | Naveed Khan, | Graham Temple, | Victoria Hough, | John Greenman,

Affiliations: Department of Biological Sciences, Hardy Building, and | Academic Surgical Unit, Wolfson Building, The University of Hull, Hull HU6 7RX, UK | PHL, Hull Royal Infirmary, Hull HU3 2JZ, UK

Note: [] Department of Biological Sciences, Hardy Building, The University of Hull, Hull HU6 7RX, UK. Tel.: +44 1482 465506; Fax: +44 1482 466996; E-mail: T.A.Paget@biosci.hull.ac.uk

Abstract: {\it Acanthamoeba} are typically identified in the laboratory using culture and microscopic observation. In this paper we describe the isolation and specificity of antibody fragments that can be used for the identification of {\it Acanthamoeba}. A phage library expressing a large repertoire (approx. 5\times 10^9) of antibody fragments was used to generate two libraries one enriched for bacteriophage that exhibit genus specific binding and the other containing bacteriophage that bind specifically to pathogenic {\it Acanthamoeba}. Individual clones were isolated on the basis of binding by ELISA, and then flow cytometry and immunofluorescence were used for further characterisation. Four monoclonal antibodies were isolated, specific for {\it Acanthamoeba} at the generic level with clone HPPG6 exhibiting the highest level of binding. Furthermore clone HPPG55 was specific for pathogenic species of {\it Acanthamoeba}.

Journal: Disease Markers, vol. 16, no. 1-2, pp. 83-90, 2000