title: Novel hapten synthesis for antibody production and development of an enzyme-linked immunosorbent assay for determination of furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ).
abstract: A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing haptens. The ability of these haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
authors.affiliation: Key Laboratory of Plant Resources Conservation and Sustainable Utilization, South China Botanical Garden, Chinese Academy of Sciences, No.723, Xingke Road, Tianhe District, Guangzhou 510650, China.
authors.forename: Zhen-Lin
authors.initials: ZL
authors.lastname: Xu
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Shen
authors.forename: Yuan-Ming
authors.initials: YM
authors.lastname: Sun
authors.forename: Katrina
authors.initials: K
authors.lastname: Campbell
authors.forename: Yuan-Xin
authors.initials: YX
authors.lastname: Tian
authors.forename: Shi-Wei
authors.initials: SW
authors.lastname: Zhang
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Lei
authors.forename: Yue-Ming
authors.initials: YM
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D002645:Chickens / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005399:Fishes / Q000378:metabolism; D006241:Haptens / Q000737:chemistry* / Q000276:immunology; D007114:Immunization; D018796:Immunoconjugates; D007074:Immunoglobulin G / Q000276:immunology; D008297:Male; D008958:Models, Molecular; D009025:Morpholines / Q000032:analysis*; D009581:Nitrofurans / Q000378:metabolism*; D023303:Oxazolidinones / Q000032:analysis* / Q000378:metabolism*; D033561:Penaeidae / Q000378:metabolism; D011817:Rabbits; D015203:Reproducibility of Results; D013552:Swine / Q000378:metabolism
chemical_list: C504349:3-amino-5-morpholinomethyl-2-oxazolidinone; D006241:Haptens; D018796:Immunoconjugates; D007074:Immunoglobulin G; D009025:Morpholines; D009581:Nitrofurans; D023303:Oxazolidinones; C007795:furaltadon