title: Generation of a highly reactive chicken-derived single-chain variable fragment against Fusarium verticillioides by phage display.
abstract: Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from entering into food/feed products. Chicken-derived single-chain variable fragments (scFvs) against cell wall-bound proteins from F. verticillioides were isolated from an immunocompetent phage display library. Comparative phage enzyme-linked immunosorbant assays (ELISAs) and sequencing analyses identified four different scFv antibodies with high sensitivity. Soluble antibody ELISAs identified two highly sensitive scFv antibodies, FvCA3 and FvCA4, with the latter being slightly more sensitive. Three-dimensional modeling revealed that the FvCA4 may hold a better overall structure with CDRH3, CDRL1 and CDRL3 centered in the core region of antibody surface compared with that of other scFvs. Immunofluorescence labeling revealed that the binding of FvCA4 antibody was localized to the cell walls of conidiospores and hyphae of F. verticillioides, confirming the specificity of this antibody for a surface target. This scFv antibody was able to detect the fungal mycelium as low as 10(-2) μg/mL and contaminating mycelium at a quantity of 10(-2) mg/g maize. This is the first report that scFv antibodies derived from phage display have a wide application for rapid and accurate detection and monitoring of fumonisin-producing pathogens in agricultural samples.
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors: Zu-Quan
authors: ZQ
authors: Hu
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors: Jin-Long
authors: JL
authors: Liu
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors: He-Ping
authors: HP
authors: Li
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.
authors: Shu
authors: S
authors: Xing
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.
authors: Sheng
authors: S
authors: Xue
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors: Jing-Bo
authors: JB
authors: Zhang
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
authors: Jian-Hua
authors: JH
authors: Wang
authors: Department Plant Biotechnology, Fraunhofer IME, Aachen 52074, Germany.
authors: Greta
authors: G
authors: Nölke
authors: Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070, China.|College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.|National Center of Plant Gene Research (Wuhan), Wuhan 430070, China.
authors: Yu-Cai
authors: YC
authors: Liao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000941:Antigens / Q000737:chemistry; D001483:Base Sequence; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D005670:Fusarium / Q000378:metabolism*; D015723:Gene Library; D021621:Imaging, Three-Dimensional; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009183:Mycotoxins / Q000737:chemistry*; D019151:Peptide Library; D017386:Sequence Homology, Amino Acid; D057127:Single-Chain Antibodies / Q000737:chemistry*; D003313:Zea mays / Q000378:metabolism
chemical_list: D000906:Antibodies; D000941:Antigens; D009183:Mycotoxins; D019151:Peptide Library; D057127:Single-Chain Antibodies