DEVELOPMENT OF MATURE AND PRO BDNF-SPECIFIC SANDWICH ELISA SYSTEM

BDNF (Brain- D erived N eurotropic F actor) plays a critical role in the nervous system. Recent studies have shown that BDNF level in serum is related to psychiatric, neurodegenerative and other aging-related chronic diseases. The enzyme-linked immunosorbent assay (ELISA) has been widely used to measure BDNF level but this assay cannot differentiate mature BDNF from proBDNF. As the function of proBDNF is different from mature BDNF, it is necessary to establish ELISA assays specific for mature BDNF and proBDNF. Objectives: The aim of this study is to establish mature and proBDNF-specific ELISA and measure their levels in samples. Polyclonal or monoclonal antibodies against mature and proBDNF were screened by Western blot and ELISA using mature BDNF and proBDNF as antigens. Specificity was tested by competition ELISA and cross-reactivity with other neruotrophins. mBDNF specific ELISA: no cross-reactivity with NT-3, NT-4, NGF; 0.99% cross-reactivity with hrproBDNF(CHO, R&D); intra-assay; 1.2 – 7.7 %C; inter-assay; 1.90– 5.64 %CV; recovery rate (human serum); 96.45-96.22 % at 1:160 dilution; LDL (low detection limit); 10-20pg/ml. proBDNF ELISA: 0.88 – 1.3 % cross-reactivity with mBDNF. HAP1KO: lower proBDNF level than WT. BACE1KO: higher mBDNF level than WT. Mature and proBDNF specific ELISA were setup. These ELISAs will be a useful method to measure mature and proBDNF level in various samples.