Using the local immune response from the natural buffalo host to generate an antibody fragment library that binds the early larval stages of Schistosoma japonicum
Christopher G Hosking, Patrick Driguez, Hamish EG McWilliam, Leodevico L Ilag, Simon Gladman, Yuesheng Li, David Piedrafita, Donald P McManus, Els NT Meeusen, Michael J de Veer
INTERNATIONAL JOURNAL FOR PARASITOLOGY | ELSEVIER SCI LTD | Published : 2015
Antibodies isolated from the local draining inguinal lymph node of field exposed-water buffaloes following challenge with Schistosoma japonicum cercariae showed high reactivity towards S. japonicum antigen preparations and bound specifically to formaldehyde-fixed S. japonicum schistosomules. Using this specific local immune response we produced a series of single-chain antibody Fv domain libraries from the same lymph nodes. Removal of phage that cross reacted with epitopes on adult parasites yielded a single-chain antibody Fv domain-phage library that specifically bound to whole formaldehyde-fixed and live S. japonicum schistosomules. DNA sequencing indicated clear enrichment of the single-c..View full abstract
The authors wish to thank Dr David Reser from Monash University, Australia and the Australian Cancer Research Foundation for Comprehensive Biomedical Imaging at QIMR Berghofer Medical Research Institute, Australia for their assistance with the fluorescent microscopy, particularly using the Zeiss 780-NLM confocal microscope. We would also like to thank Prof. Andreas Plueckthun at the University of Zurich, Switzerland for kindly supplying the pAK100 phagemid vector, Dr Melanie Neeland for reading and improving the manuscript, Dieter Bulach at Victorian Life Sciences Computation Initiative, Australia and David Powell at the Monash Bioinformatics Platform, Australia for their help analysing the Ion Torrent sequencing data and The Australian Society for Parasitology (ASP) for travel funds. DPM is a National Health Medical Research Centre, Australia, Research Senior Principal Research Fellow and a Senior Scientist at QIMR Berghofer Medical Research Institute.