Selection and Characterization of CSFV-Specific Single-Domain Antibodies and Their Application along with Immunomagnetic Nanobeads and Quantum Dots
Shunli Yang , Li Yuan , Youjun Shang , Jinyan Wu , Xiangtao Liu , Jie Zhang , Zygmunt Pejsak , Katarzyna Podgórska , Katarzyna Stepniewska , Muhammad Umar Zafar Khan , Jianping Cai , Shuanghui Yin
AbstractOutbreak of classical swine fever (CSF) results in high mortality and thus causes severe economic losses in the swine industry. Single-domain antibody (sdAb) is the smallest antigen-binding molecule derived from camelid heavy-chain antibodies and has the potential to be used as a molecular probe for detection of CSF virus (CSFV). In this study, two sdAb fragments against the E2 antigen of CSFV were obtained, expressedin vitro. The functional characteristics analysis indicated that the recombinant sdAbE2-1 and sdAbE2-2 have excellent binding activity, specificity, and high affinity with equilibrium constant value of 3.34 × 10−7and 1.35 × 10−8 M to E2 protein. Then, sdAbE2s were conjugated with quantum dots (QD)/AF488 to synthesize two molecular probes for imaging CSFV distribution in cells. The sdAbE2-1 was also labeled with carboxyl-magnetic beads to construct immunomagnetic nanobeads (IMNBs) able to capture CSFV virions and recombinant E2 protein. QD/AF455-sdAbE2s probes colocalised with CSFV virions in swine testis cells, and IMNBs were used as a detection template and proved to bind specifically with CSFV virions and E2 protein. The selected sdAb fragments and sdAb-based molecular probes may be used for the rapid identification of CSFV during field outbreaks and for research on CSFV and host interactions.
|Journal series||BioMed Research International, [Journal of Biomedicine and Biotechnology], ISSN 1110-7243, e-ISSN 2314-6141, [1110-7251, 2314-6133, 2314-6141], (N/A 70 pkt, Not active)|
|Publication size in sheets||0.5|
|ASJC Classification||; ; ; ; ; ; ;|
|Publication indicators||: 2016 = 0.875; : 2018 = 2.197 (2) - 2018=2.885 (5)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.