Parallel selection of multiple anti-infectome Nanobodies without access to purified antigens.

TitleParallel selection of multiple anti-infectome Nanobodies without access to purified antigens.
Publication TypeJournal Article
Year of Publication2008
AuthorsSaerens, D., Stijlemans B., Baral T. Nath, Thi G. Thanh Nguy, Wernery U., Magez S., De Baetselier P., Muyldermans S., and Conrath K.
JournalJ Immunol Methods
Volume329
Issue1-2
Pagination138-50
Date Published2008 Jan 1
Type of Articlenanobody
ISSN0022-1759
KeywordsAmino Acid Sequence, Animals, Antibodies, Protozoan, Antibody Specificity, Camels, Cell Separation, Cloning, Molecular, Cross Reactions, Enzyme-Linked Immunosorbent Assay, Feasibility Studies, Flow Cytometry, Gene Library, Immunoglobulin Fab Fragments, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Nanotechnology, Trypanosoma, Variant Surface Glycoproteins, Trypanosoma
Abstract

A strategy was developed to isolate Nanobodies, camelid-derived single-domain antibody fragments, against the parasite infectome without a priori knowledge of the antigens nor having access to the purified antigens. From a dromedary, infected with T. evansi, we cloned a pool of Nanobodies and selected after phage display 16 different Nanobodies specific for a single antigen, i.e. variant surface glycoprotein of T. evansi. Moreover 14 Nanobodies were isolated by panning on different total parasite lysates. Thus, this anti-infectome experiment generated Nanobodies, monospecific for one Trypanosoma species, whereas others were pan-reactive to various Trypanosoma species. Several Nanobodies could label specifically the coat of a set of Trypanozoon species. The recognized target(s) are present in GPI-linked membrane fractions of bloodstream- and fly-form parasites. Due to the omnipresence of these targets on different parasite species and forms, these antibody fragments are a valuable source for validation of novel, not yet identified targets to design new diagnostics and therapeutics.

DOI10.1016/j.jim.2007.10.005
Alternate JournalJ. Immunol. Methods
PubMed ID17996887