GALLO CALDERON marina beatriz
congresos y reuniones científicas
OBTENTION OF SPECIFIC SINGLE DOMAIN ANTIBODIES DERIVED FROM CAMELIDS (VHH OR NANOBODIES) AGAINST PLASMA BIOMARKERS OF ALZHEIMER?S DISEASE FOR DIAGNOSTIC APPLICATION.
JOAQUIN BOZZO; GALLO CALDERON, MARINA; MATTION, NORA; VANINA GRIPPO
Mar del Plata
Congreso; LXI REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA (SAIC) LXIV REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INMUNOLOGÍA (SAI) XLVIII REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE FARMACOLOGÍA EXPERIMENTAL (SAFE) VII REUNIÓN ANUAL DE LA SOCIEDAD; 2016
Alzheimer´s disease A& affects 0ervous Central System beforeclinical symptoms are fully expressed. Currently A& diagnosisdepends on clinical evaluations, and there is no reliable earlydiagnostic test. &irect measure of plasma biomarMers for A&, suchas alphaantitrypsin AAT and alphamacroglobulin A/,could lead to an earlier and more sensitive diagnosis. 0oYadays,single domain antibodies derived from camelids 8HH or nanobodies represent a very useful tool Yith unique characteristics,maMing 8HH more suitable for sensitive and specific diagnosis.For this reason, Ye aimed to obtain and produce specific 8HHagainst bloodbased protein biomarMers for A&, AAT and A/,Yith potential application in the development of a simple and earlydiagnostic test.+n this YorM, tYo llamas Yere immunized Yith A/and A1AT proteins obtaining a high humoral response in both animals.TYo 8HH libraries Yere constructed starting from peripheralblood lymphocytes from both llamas. Library quality controls Yereperformed to corroborate fulllength 8HH insert and repertoire variability.A high percentage of fulllength clones . and and at least different fingerprinting profiles Yere determined foreach library. Then, phages that expressed in its surface specific8HH against both biomarMers Yere selected by 2hage &isplaymethodology. After tYoround selection, specific reactivity of and selected clones for A/ and AAT respectively, Yasconfirmed by EL+SA. Four most reactive nanobodiesYere subcloned into pHE0 vector and expressed as soluble protein in E.coli. 8HH expression Yas confirmed by 9estern $lot detecting aunique band of approximately M& as expected. Finally, specificbiomarMer recognition by expressed 8HH Yas confirmed by EL+SAand 9estern $lot. +n conclusion, specific 0anobodiesagainstbloodbased protein biomarMers of A&, A/ and AAT, Yereobtained, expressed and preliminarily characterized for potentialapplication in AD diagnosis.