Mycotoxins, which are toxic, carcinogenic, and/or teratogenic, have posed a threat to food safety and public health. Sensitive and effective determination technologies for mycotoxin surveillance are required. Immunoassays have been regarded as useful supplements to chromatographic techniques. However, conventional antibodies involved in immunoassays are difficult to be expressed recombinantly and are susceptible to harsh environments. Nanobodies (or VHH antibodies) are antigen-binding sites of the heavy-chain antibodies produced from Camelidae. They are found to be expressed easily in prokaryotic or eukaryotic expression systems, more robust in extreme conditions, and facile to be used as surrogates for artificial antigens. These properties make them the promising and environmentally friendly immunoreagents in the next generation of immunoassays. This review briefly describes the latest developments in the area of nanobodies used in mycotoxin detection. Moreover, by integrating the introduction of the principle of nanobodies production and the critical assessment of their performance, this paper also proposes the prospect of nanobodies in the field of food safety in the foreseeable future.

To develop a functional alternative hepatocyte model for primary human hepatocytes (PHH) with proliferative property, essential drug metabolic and transporter functions, proliferating human hepatocytes (ProliHH) expanded from PHH were fully characterized invitro. Herein, ProliHH generated from multiple PHH donors could be expanded more than 200 folds within four passages, maintained their metabolic or transporter capacities partially, and further were able to regain of the mature hepatic property after three-dimensional (3D) culture. Particularly, the downregulated mRNA expression and function of three major CYP450 enzymes (CYP1A2, CYP2B6, and CYP3A4) in the proliferating process (ProliHH-P) could be recovered by 3D culture. The metabolic variabilities across different PHH donors could be inherited to their matured ProliHH (ProliHH-M). The intrinsic clearances (CL_int) of seven major CYP450 enzymes in ProliHH-M were correlated well (r = 0.87) with that in PHH. Also, bile canaliculi structures could be observed in sandwich cultured ProliHH (SC-ProliHH), and the biliary excretion index (BEI) of four probe compounds (CLF, CDF, d8-TCA and rosuvastatin) in SC-ProliHH (>10%) were comparable to SC-PHH. More importantly, both ProliHH-P and ProliHH-M could be used to evaluate hepatotoxicity, and ProliHH-M were more suitable for transporter mediated drug hepatotoxicity evaluation. Therefore, these findings demonstrated that the 3D and sandwich culture system could be utilized to recover and maintain metabolic and transporter functions in ProliHH for clearance prediction and cholestasis risk assessment, respectively. ProliHH could be a promising substitute for PHH in drug metabolism, transport and hepatotoxicity screening. Significance Statement We report the first demonstration that drug metabolic capacities and efflux transporter functions of ProliHH, as well as their potential in toxicity assessments. The metabolic variability in different primary human hepatocytes (PHH) donors could be inherited by their derived matured ProliHH. ProliHH could form canalicular networks in sandwich culture. These results make it feasible for ProliHH to support drug candidate screening in hepatic metabolism, disposition and toxicity.

TGIF1 is a transcriptional repressor playing crucial roles in human development and function and is associated with holoprosencephaly and various cancers. TGIF1-directed transcriptional repression of specific genes depends on the recruitment of corepressor SIN3A. However, to date, the exact region of TGIF1 binding to SIN3A was not clear, and the structural basis for the binding was unknown. Here, we demonstrate that TGIF1 utilizes a C-terminal domain (termed as SIN3A-interacting domain, SID) to bind with SIN3A PAH2. The TGIF1 SID adopts a disordered structure at the apo state but forms an amphipathic helix binding into the hydrophobic cleft of SIN3A PAH2 through the nonpolar side at the holo state. Residues F379, L382 and V383 of TGIF1 buried in the hydrophobic core of the complex are critical for the binding. Moreover, homodimerization of TGIF1 through the SID and key residues of F379, L382 and V383 was evidenced, which suggests a dual role of TGIF1 SID and a correlation between dimerization and SIN3A-PAH2 binding. This study provides a structural insight into the binding of TGIF1 with SIN3A, improves the knowledge of the structure–function relationship of TGIF1 and its homologs and will help in recognizing an undiscovered SIN3A-PAH2 binder and developing a peptide inhibitor for cancer treatment.