title: Characterization of Asia 1 sdAb from camels bactrianus (C. bactrianus) and conjugation with quantum dots for imaging FMDV in BHK-21 cells.
abstract: Foot-and-mouth disease (FMD), caused by FMD virus (FMDV), is a highly contagious viral disease affecting cloven-hoofed animals. Camelids have a unique immunoglobulin profile, with the smallest functional heavy-chain antibodies (sdAb or VHH) naturally devoid of light chains with antigen-binding capacity. We screened and characterized five sdAbs against FMDV by immunized library from C. bactrianus with Asia 1 virus-like particles (VLPs). Three of five recombinant sdAbs were stably expressed in E.coli, remained highly soluble, and were serotype-specific for VP1 protein of FMDV Asia 1 by ELISA. These failed to completely neutralize the Asia 1 virus. According to the KD value of binding affinity to three sdAbs, which ranged from 0.44 to 0.71 nm by SPR, sdAb-C6 was selected and conjugated with Zn/CdSe quantum dots (QDs) to form a QDs-C6 probe, which was used to trace and image the subcellular location of FMDV in BHK-21 cells. The results show that FMD virions were observed from 3 h.p.i., and most of virions were distributed on one side of the nucleus in the cytoplasm. We demonstrate the utility of sdAbs as functionalized QDs are powerful tools for FMDV research.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
authors.forename: Shuanghui
authors.initials: S
authors.lastname: Yin
authors.forename: Shunli
authors.initials: S
authors.lastname: Yang
authors.forename: Youjun
authors.initials: Y
authors.lastname: Shang
authors.forename: Shiqi
authors.initials: S
authors.lastname: Sun
authors.forename: Guangqing
authors.initials: G
authors.lastname: Zhou
authors.forename: Ye
authors.initials: Y
authors.lastname: Jin
authors.forename: Hong
authors.initials: H
authors.lastname: Tian
authors.forename: Jinyan
authors.initials: J
authors.lastname: Wu
authors.forename: Xiangtao
authors.initials: X
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D019187:Cadmium Compounds / Q000737:chemistry; D002162:Camelus / Q000276:immunology / Q000821:virology*; D002460:Cell Line; D006224:Cricetinae; D005260:Female; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology* / Q000302:isolation & purification* / Q000502:physiology; D042541:Intracellular Space / Q000821:virology; D008297:Male; D057054:Molecular Imaging / Q000379:methods*; D008969:Molecular Sequence Data; D045663:Quantum Dots; D018036:Selenium Compounds / Q000737:chemistry; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012995:Solubility; D013440:Sulfides / Q000737:chemistry; D017967:Zinc Compounds / Q000737:chemistry
chemical_list: D000914:Antibodies, Viral; D019187:Cadmium Compounds; D018036:Selenium Compounds; D057127:Single-Chain Antibodies; D013440:Sulfides; D017967:Zinc Compounds; C058667:cadmium selenide; C031238:zinc sulfide