Synthesis of site-heterologous haptens for high-affinity anti-pyraclostrobin antibody generation

Josep V. Mercader, Consuelo Agulló, Antonio Abad-Somovilla, Antonio Abad-Fuentes
2011 Organic and biomolecular chemistry  
The design and synthesis of functional chemical derivatives of small organic molecules is usually a key step for the intricate production of a variety of bioconjugates. In this respect, the derivatization site at which the spacer arm is introduced in immunizing conjugates constitutes a highly critical parameter for the generation of high-affinity and selective antibodies. However, due to the usual complexity of the required synthetic procedures, the appropriate comparison of alternative
more » ... alternative tethering 10 positions has often been neglected. In the present study, meticulous strategies were followed to prepare synthetic derivatives of pyraclostrobin with the same linkers located at diverse rationally-chosen sites. Activity appraisal of antibodies and bioconjugates was carried out by bidimensional competitive direct and indirect immunoassays, and a superior performance of two of the three synthesized haptens was found. Finally, a detailed analysis of the conformations of the target molecule and the synthesized haptens in aqueous solution was done using computer assisted molecular modeling techniques. This study suggested 15 that the lower titers and affinities of one set of antibodies are most probably due to conformational effects of the spacer arm in the immunizing bioconjugate. 65 65 This journal is © The Royal Society of Chemistry [year] Journal Name, [year], [vol], 00-00 | 3 115 Notes and references
doi:10.1039/c0ob00686f pmid:21225057 fatcat:heriswgsingdvjqyl3jknjuvbe