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

Abstract

Outbreak 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.
Author Shunli Yang
Shunli Yang,,
-
, Li Yuan
Li Yuan,,
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, Youjun Shang
Youjun Shang,,
-
, Jinyan Wu
Jinyan Wu,,
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, Xiangtao Liu
Xiangtao Liu,,
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, Jie Zhang
Jie Zhang,,
-
, Zygmunt Pejsak (VET)
Zygmunt Pejsak,,
- University Centre for Veterinary Medicine UJ-UR
, Katarzyna Podgórska
Katarzyna Podgórska,,
-
, Katarzyna Stepniewska
Katarzyna Stepniewska,,
-
, Muhammad Umar Zafar Khan
Muhammad Umar Zafar Khan,,
-
et al.`
Journal seriesBioMed 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)
Issue year2020
Vol2020
Pages1-11
Publication size in sheets0.5
Article number3201630
ASJC Classification2700 General Medicine; 1300 General Biochemistry, Genetics and Molecular Biology; 1305 Biotechnology; 1311 Genetics; 1312 Molecular Biology; 1313 Molecular Medicine; 2400 General Immunology and Microbiology; 2307 Health, Toxicology and Mutagenesis
DOIDOI:10.1155/2020/3201630
Languageen angielski
File
3201630.pdf 4,07 MB
Score (nominal)70
Score sourcejournalList
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2016 = 0.875; WoS Impact Factor: 2018 = 2.197 (2) - 2018=2.885 (5)
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