Synucleinopathies are conditions that remain with no available effective treatments thus far. Immunotherapy is a possible path to fight against such pathologies by inducing antibodies against alpha-synuclein (α-Syn), which could induce the clearance of its pathologic form. Looking to develop a new low-cost, effective vaccine against synucleinopathies; we have designed a chimeric plant-made antigen comprising the subunit B of the enterotoxin from enterotoxigenic E. coli and three B cell epitopes from α-Syn, which is named LTB-Syn. In the present study, LTB-Syn was produced in carrot cell lines as appropriate platform for the formulation of oral vaccines not requiring purification. The development of transgenic carrot cell lines took 8 months and the LTB-Syn yield reached 2.3 μg/g dry biomass. The antigen encapsulated in lyophilized carrot cells was highly stable at room temperature over a six-month period and upon heating at 50 °C for 2 h. Moreover, LTB-Syn was able to prime immune responses that, in combination with parenteral boosting using an OVA-Syn conjugate, induced significant humoral resposes in mice. Thus the carrot-made oral LTB-Syn vaccine is a promising candidate that deserves further analyses to advance in its preclinical evaluation.,Copyright © 2019 Elsevier B.V. All rights reserved.