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- Title
Characterization of Asia 1 sdAb from Camels Bactrianus (<i>C. bactrianus</i>) and Conjugation with Quantum Dots for Imaging FMDV in BHK-21 Cells
- Authors
Yin, Shuanghui; Yang, Shunli; Shang, Youjun; Sun, Shiqi; Zhou, Guangqing; Jin, Ye; Tian, Hong; Wu, Jinyan; Liu, Xiangtao
- 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.
- Subjects
ASIA; CAMELS; ZOOLOGICAL microtechnique; QUANTUM dot devices; IMAGING systems; FOOT &; mouth disease; IMMUNOGLOBULINS; ANTIGENS; PROTEIN binding
- Publication
PLoS ONE, 2013, Vol 8, Issue 5, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0063500