title: An immunogen synthesis strategy for the development of specific anti-deoxynivalenol monoclonal antibodies.
abstract: An immunogen synthesis strategy was designed to develop anti-deoxynivalenol (DON) monoclonal antibodies with low cross-reactivity against structurally similar trichothecenes. A total of eight different DON immunogens were synthesised, differing in the type and position of the linker on the DON molecule. After immunisation, antisera from mice immunised with different DON immunogens were checked for the presence of relevant antibodies. Then, both homologous and heterologous enzyme-linked immunosorbent assays (ELISAs) were performed for hybridoma screening. Finally, three monoclonal antibodies against DON and its analogues were generated. In addition, monoclonal antibody 13H1 could recognise DON and its analogues in the order of HT-2 toxin > 15-acetyldeoxynivalenol (15-ADON) > DON, with IC₅₀ ranging from 1.14 to 2.13 µg ml⁻¹. Another monoclonal antibody 10H10 manifested relatively close sensitivities to DON, 3-acetyldeoxynivalenol (3-ADON) and 15-ADON, with IC₅₀ values of 22, 15 and 34 ng ml⁻¹, respectively. Using an indirect ELISA format decreases the 10H10 sensitivity to 15-ADON with 92%. A third monoclonal antibody 2A9 showed to be very specific and sensitive to 3-ADON, with IC₅₀ of 0.38 ng ml⁻¹. Using both 2A9 and 10H10 monoclonal antibodies allows determining sole DON contamination.
authors.affiliation: a Laboratory of Food Analysis , Ghent University , Ghent , Belgium.
authors.forename: Melanie
authors.initials: M
authors.lastname: Sanders
authors.forename: Yirong
authors.initials: Y
authors.lastname: Guo
authors.forename: Abhishek
authors.initials: A
authors.lastname: Iyer
authors.forename: Yara Ruiz
authors.initials: YR
authors.lastname: García
authors.forename: Anastasia
authors.initials: A
authors.lastname: Galvita
authors.forename: Arne
authors.initials: A
authors.lastname: Heyerick
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.forename: Martijn D P
authors.initials: MD
authors.lastname: Risseeuw
authors.forename: Serge
authors.initials: S
authors.lastname: Van Calenbergh
authors.forename: Marc
authors.initials: M
authors.lastname: Bracke
authors.forename: Sergei
authors.initials: S
authors.lastname: Eremin
authors.forename: Annemieke
authors.initials: A
authors.lastname: Madder
authors.forename: Sarah
authors.initials: S
authors.lastname: De Saeger
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism*; D000915:Antibody Affinity; D000918:Antibody Specificity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005506:Food Contamination; D005513:Food Inspection / Q000379:methods; D006825:Hybridomas / Q000378:metabolism; D007700:Kinetics; D008807:Mice, Inbred BALB C; D018448:Models, Immunological; D008958:Models, Molecular; D015394:Molecular Structure; D011042:Poisons / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D014255:Trichothecenes / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000737:chemistry; D014614:Vaccines, Synthetic / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D011042:Poisons; D014255:Trichothecenes; D014614:Vaccines, Synthetic; C007262:deoxynivalenol
keywords: ELISA; cross-reactivity; deoxynivalenol; immunogens; monoclonal antibodies