Parallel selection of multiple anti-infectome Nanobodies without access to purified antigens.
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.
Saerens
Dirk
D
Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussels, Pleinlaan 2, B-1050 Brussels, Belgium. dsaerens@vub.ac.be
Stijlemans
Benoît
B
Baral
Toya Nath
TN
Nguyen Thi
Giang Thanh
GT
Wernery
Ulrich
U
Magez
Stefan
S
De Baetselier
Patrick
P
Muyldermans
Serge
S
Conrath
Katja
K
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D000918:Antibody Specificity; D002162:Camelus; D002469:Cell Separation; D003001:Cloning, Molecular; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D005434:Flow Cytometry; D015723:Gene Library; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D036103:Nanotechnology; D014345:Trypanosoma / Q000276:immunology*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000097:blood / Q000276:immunology*
D000913:Antibodies, Protozoan; D007140:Immunoglobulin Fab Fragments; D014643:Variant Surface Glycoproteins, Trypanosoma