< Back to previous page
Project
Nanobody-based proximity ligation assay for human toxocariasis. (Nanobody)
In Human toxocariasis, severe disease may issue from infection with one single Toxocara larva. Diagnosis is difficult because the parasite does not replicate in the host and is located in tissues. Serological tests remain the least invasive and most sensitive approach to detect parasite antigens (Ags), but have thus far lacked sensitivity and specificity. Novel approaches are urgently needed to deliver tests of clinical and epidemiological relevance.
Our strategy exploits the advantages of two powerful techniques to address these needs: Nanobodies (Nbs) and the Proximity Ligation Assay (PLA).
Specificity will be addressed by generating Nbs as Agn-detection reagents. Nbs are recombinant, camel derived antibodies (Abs) recognizing Agn with high affinity and specificty. Their unique flexible in vitro selection procedure allows the generation of Nbs which unambiguously detect larval Agn (ie in cysticercosis). An added benefit of Nbs is that their small size leads to the recognition of epitopes on the Agn that are not antigenic for classical Abs so that the presence of host Abs will not compete with the Agn detection by Nbs.
Improved sensitivity will be attained by coupling Nbs to the PLA, a technique converting protein recognition events (Nbs to Agn) into a DNA readout. Detection of Agn by DNAconjugated Nbs leads to the ligation of unique DNA sequences, amplified by PCR to surpass the sensitivity of conventional protein detection methods.
Our strategy exploits the advantages of two powerful techniques to address these needs: Nanobodies (Nbs) and the Proximity Ligation Assay (PLA).
Specificity will be addressed by generating Nbs as Agn-detection reagents. Nbs are recombinant, camel derived antibodies (Abs) recognizing Agn with high affinity and specificty. Their unique flexible in vitro selection procedure allows the generation of Nbs which unambiguously detect larval Agn (ie in cysticercosis). An added benefit of Nbs is that their small size leads to the recognition of epitopes on the Agn that are not antigenic for classical Abs so that the presence of host Abs will not compete with the Agn detection by Nbs.
Improved sensitivity will be attained by coupling Nbs to the PLA, a technique converting protein recognition events (Nbs to Agn) into a DNA readout. Detection of Agn by DNAconjugated Nbs leads to the ligation of unique DNA sequences, amplified by PCR to surpass the sensitivity of conventional protein detection methods.
Date:1 Jan 2013 → 31 Dec 2018
Project type:PhD project