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Disease Markers
Volume 16 (2000), Issue 1-2, Pages 83-90
http://dx.doi.org/10.1155/2000/984538

Use of Phage Antibodies to Distinguish Closely Related Species of Protozoan Parasites

Timothy Paget,1 Naveed Khan,1 Graham Temple,3 Victoria Hough,2 and John Greenman2

1Department of Biological Sciences, Hardy Building, The University of Hull, Hull HU6 7RX, UK
2Academic Surgical Unit, Wolfson Building, The University of Hull, Hull HU6 7RX, UK
3PHL, Hull Royal Infirmary, Hull HU3 2JZ, UK

Received 12 March 2001; Accepted 12 March 2001

Copyright © 2000 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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 Acanthamoeba. A phage library expressing a large repertoire (approx. 5×109) 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 Acanthamoeba. Individual clones were isolated on the basis of binding by ELISA, and then flowcytometry and immunofluorescence were used for further characterisation. Four monoclonal antibodies were isolated, specific for Acanthamoeba at the generic level with clone HPPG6 exhibiting the highest level of binding. Furthermore clone HPPG55 was specific for pathogenic species of Acanthamoeba.