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Immunoassay relies on antibodies, but traditional antibodies such as monoclonal antibody (mAb) require animal immunization and complex procedures. Single-chain variable fragment (scFv) becomes a potential alternative to mAb with advantages of the low cost, rapid and easy prepared. In the present study, we prepared scFvs against dihydroartemisinin (DHA) based on E. coli and HEK293T cell expression system, named MBP-scFv and scFv-Fc, respectively. Their properties were compared with the parent mAb. The calculated affinity constants of mAb, MBP-scFv and scFv-Fc were 2.1 × 10 8  L mol -1 , 2.2 × 10 7  L mol -1 and 1.6 × 10 8  L mol -1 , respectively. The half inhibitory concentration (IC 50 ) of mAb, MBP-scFv and scFv-Fc were 1.16 ng mL -1 , 2.15 ng mL -1 and 6.57 ng mL -1 , respectively. Both the scFv showed less sensitive than the mAb based on the IC 50 . The cross-reactivities of MBP-scFv for artemisinin and artesunate exhibited similarities to the mAb, yet the cross-reactivities of scFv-Fc for these compounds exceeded those of the mAb significantly. The stability of the scFvs was ascertained to be maintained for over 5 days at room temperature, and for more than a month at both 4 °C and - 20 °C. After that, the indirect competitive enzyme-linked immunosorbent assays (icELISAs) based on the scFv from E. coli were used to detect the DHA content in eight drug samples, and the result was consistent with ultra-performance liquid chromatography simultaneously. Although scFv can be used for quantitative determination of drugs, but it still cannot completely replace mAb in immunoassay without evolution and modification., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The rhizobium-legume symbiosis system is critical for nitrogen-cycle balance in agriculture. However, the potential effects of carbon nanomaterials (CNMs) on this system remain largely unknown. Herein, we studied the effects of four carbon-based materials (activated carbon (AC), single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs) and graphene oxide (GO)) on the rhizobium-legume symbiosis system consisting of Lotus japonicus and Mesorhizobium loti MAFF303099. Under non-symbiotic conditions, the bacterial growth and root development of plants were both clearly inhibited by SWCNTs and GO, while the elongation of plant stems was enhanced by MWCNTs to a certain degree. More importantly, only MWCNTs could increase the number of nodules and enhance the activity of nitrogenase in the rhizobium-plant interaction. Further analyses showed that the average number of nodules in plants treated with 100 μg mL -1 MWCNTs was significantly increased by 39% at 14 days post inoculation (dpi) and by 41% at 28 dpi. Meanwhile, the biological nitrogen fixation of the nodules was promoted by more than 10% under 100 μg mL -1 MWCNT treatment, which enhanced the above- and below-ground fresh biomass by 14% and 25% respectively at 28 dpi. Transmission electron microscopy images further indicated that MWCNTs penetrated the cell wall, and pierced through the cell membrane to be transmitted into the cytoplasm. In addition, gene expression analysis showed that the promotion of nodulation by MWCNTs was correlated with the up-regulation of certain genes involved in this signaling pathway. In particular, the expression of NIN, a crucial gene regulating the development of nodules, was significantly elevated 2-fold by MWCNTs at an early stage of nodulation. These findings are expected to facilitate the understanding and future utilization of MWCNTs in agriculture.

To understand the interaction mechanism between graphene oxide (GO) and typical phytopathogens, a particular investigation was conducted about the antimicrobial activity of GO against two bacterial pathogens (P. syringae and X. campestris pv. undulosa) and two fungal pathogens (F. graminearum and F. oxysporum). The results showed that GO had a powerful effect on the reproduction of all four pathogens (killed nearly 90% of the bacteria and repressed 80% macroconidia germination along with partial cell swelling and lysis at 500 μg mL(-1)). A mutual mechanism is proposed in this work that GO intertwinds the bacteria and fungal spores with a wide range of aggregated graphene oxide sheets, resulting in the local perturbation of their cell membrane and inducing the decrease of the bacterial membrane potential and the leakage of electrolytes of fungal spores. It is likely that GO interacts with the pathogens by mechanically wrapping and locally damaging the cell membrane and finally causing cell lysis, which may be one of the major toxicity actions of GO against phytopathogens. The antibacterial mode proposed in this study suggests that the GO may possess antibacterial activity against more multi-resistant bacterial and fungal phytopathogens, and provides useful information about the application of GO in resisting crop diseases.