Title | Parallel selection of multiple anti-infectome Nanobodies without access to purified antigens. |
Publication Type | Journal Article |
Year of Publication | 2008 |
Authors | Saerens, D., Stijlemans B., Baral T. Nath, Thi G. Thanh Nguy, Wernery U., Magez S., De Baetselier P., Muyldermans S., and Conrath K. |
Journal | J Immunol Methods |
Volume | 329 |
Issue | 1-2 |
Pagination | 138-50 |
Date Published | 2008 Jan 1 |
Type of Article | nanobody |
ISSN | 0022-1759 |
Keywords | Amino 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. |
DOI | 10.1016/j.jim.2007.10.005 |
Alternate Journal | J. Immunol. Methods |
PubMed ID | 17996887 |
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