A Novel Full-length IgG Recombinant Antibody Highly Specific to Clothianidin and Its Application in Immunochromatographic Assay

The toxicity of clothianidin to non-target organisms has gradually attracted world-wide attention. It is essential to develop reliable methods for the on-site detection of clothianidin residue. In this study, analogue-based heterologous ic-ELISAs were designed to rapidly screen desirable hybridomas, which could be used for the construction of recombinant antibodies (RAbs) against clothianidin. Based on the antibody variable region genes, two full-length IgG RAbs (1F7-RAb and 5C3-RAb) were produced by the mammalian cell expression system. The performance of the two RAbs was characterized and compared by heterologous ic-ELISAs and non-competitive surface plasmon resonance (SPR) assays. Using heterologous ic-ELISAs, the 1F7-RAb exhibited highly specific and sensitive recognition to clothianidin with an IC50 of 4.62 mu g/L whereas the 5C3-RAb could bind to both clothianidin and dinotefuran. The results of the non-competitive SPR assay further verified that the 1F7-RAb had a higher specificity and affinity to clothianidin than the 5C3-RAb. Finally, a gold immunochromatographic assay based on the novel antibody, 1F7-RAb, was developed for rapid detection of clothianidin with high sensitivity (visual detection limit of 2.5 mu g/L), specificity, and good reproducibility, which can be used as an effective supervision tool for clothianidin residue in agricultural and environmental samples.

Automated summary

The development of reliable methods for on-site detection of clothianidin residue is essential due to increasing concern about its toxicity. In this study, analogue-based heterologous ic-ELISAs were used to screen desirable hybridomas for the production of recombinant antibodies (RAbs) against clothianidin. The performance of two full-length IgG RAbs (1F7-RAb and 5C3-RAb) was characterized and compared using heterologous ic-ELISAs and non-competitive surface plasmon resonance (SPR) assays. The 1F7-RAb exhibited highly specific and sensitive recognition to clothianidin and was successfully used to develop a gold immunochromatographic assay for rapid detection of clothianidin.