title: A Plant-Based Transient Expression System for the Rapid Production of Malaria Vaccine Candidates.
abstract: There are currently no vaccines that provide sterile immunity against malaria. Various proteins from different stages of the Plasmodium falciparum life cycle have been evaluated as vaccine candidates, but none of them have fulfilled expectations. Therefore, combinations of key antigens from different stages of the parasites life cycle may be essential for the development of efficacious malaria vaccines. Following the identification of promising antigens using bioinformatics, proteomics, and/or immunological approaches, it is necessary to express, purify, and characterize these proteins and explore the potential of fusion constructs combining different antigens or antigen domains before committing to expensive and time-consuming clinical development. Here, using malaria vaccine candidates as an example, we describe how Agrobacterium tumefaciens-based transient expression in plants can be combined with a modular and flexible cloning strategy as a robust and versatile tool for the rapid production of candidate antigens during research and development.
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Boes
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Reimann
authors.affiliation: TRM Ltd, PO Box 93, York, YO43 3WE, UK.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Twyman
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany.|Institute for Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schillberg
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany. holger.spiegel@ime.fraunhofer.de.
authors.forename: Holger
authors.initials: H
authors.lastname: Spiegel
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D017780:Malaria Vaccines / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics; D013997:Time Factors; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D017780:Malaria Vaccines
keywords: Agrobacterium tumefaciens; DsRed; Gene stacking; Modular cloning; Nicotiana benthamiana; Recombinant protein production