Characterization of single-domain antibodies against Foot and Mouth Disease Virus (FMDV) serotype O from a camelid and imaging of FMDV in baby hamster kidney-21 cells with single-domain antibody-quantum dots probes

Wang, Di; Yang, Shunli; Yin, Shuanghui; Shang, Youjun; Du, Pu; Guo, Jianpeng; He, Jijun; Cai, Jianping; Liu, Xiangtao

doi:10.1186/s12917-015-0437-2

Cited by 12 publications

(10 citation statements)

References 41 publications

(46 reference statements)

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“…The conjugate of QDs and antibodies has been recognized as a powerful bioprobe and applied widely in ELISA and immunofluorescence (Goldman et al 2002;Umakoshi et al 2018). Given the excellent property of sdAb in terms of structure, stability, and production costs, sdAbs have been used successfully as an attractive alternative to conjugate with QDs for the imaging and diagnostic platforms (Sukhanova et al 2012;Wang et al 2015;Yin et al 2013). In the current study, sdAb-Mc37 showed high Kd value, conjugated with a ZnS-capped CdSe QDs.…”

Section: Discussionmentioning

confidence: 99%

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

Yang

Yin

et al. 2018

Appl Microbiol Biotechnol

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Single-domain antibody (sdAb) or nanobody possesses specific features non-accessible for conventional antibodies that make them suitable for research and biotechnological applications. Porcine epidemic diarrhea virus (PEDV) causes lethal diarrhea in piglets, resulting in great economic losses all over the world. To detect and isolate PEDV rapidly and accurately is important for the control and further research of the clinical PEDV strains. In this study, four sdAb fragments (sdAb-Mc19/29/30/37) targeting the membrane (M) protein of PEDV were selected from sdAb library that was constructed through M protein-immunized Camelus bactrianus. The selected sdAb-Mcs were solubly expressed in Escherichia coli. The functional characteristics analysis revealed that the recombinant sdAb-Mcs have excellent binding activity and specificity to M protein but have no neutralizing activity to PEDV. For further application, sdAb-Mc37 was conjugated with quantum dots to synthesize a nanoprobe for imaging PEDV in vero cells. The observed fluorescence in vero cells clearly reflects that PEDV virions can be reliably recognized and labeled by the nanoprobe. Furthermore, the sdAb-Mc29 was conjugated with superparamagnetic nanobeads to construct immunomagnetic nanobeads (IMNBs) used to isolate PEDV. One PEDV strain was successfully isolated from clinical fecal sample, suggesting IMNBs as a novel and efficient tool suitable for PEDV isolation from clinical samples. This study provided a novel application and substantiated the suitability of sdAb as a specific binder for the isolation of viruses.

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Section: Discussionmentioning

confidence: 99%

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

Yang

Yin

et al. 2018

Appl Microbiol Biotechnol

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“…QD605-Labeled sdAbE2-1 and AF488-Labeled sdAbE2-2 as Imaging Probes for CSFV in ST Cells. Activated QD 605nm (ZnS-capped CdSe) was modified with thioglycolic acid and then subjected to reaction with 1 mg of sdAbE2-1 to obtain the QD-sdAbE2-1 probe following a previously described protocol [23,24]. AF488 (Alexa Fluor 488, Wuhan Jiayuan QuantumDots Co. Wuhan, China) was conjugated with sdAb-E2-2 forming AF488-sdAbE2-2.…”

Section: Production Of Immunomagnetic Nanobeads Formentioning

confidence: 99%

Selection and Characterization of CSFV-Specific Single-Domain Antibodies and Their Application along with Immunomagnetic Nanobeads and Quantum Dots

Yang¹,

Li²,

Shang³

et al. 2020

BioMed Research International

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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, expressed in vitro. e 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 − 7 and 1.35 × 10 − 8 M to E2 protein. en, sdAbE2s were conjugated with quantum dots (QD)/AF488 to synthesize two molecular probes for imaging CSFV distribution in cells. e 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. e 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.

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“…Specific VHHs raised against FMD Asia 1 virus have also been obtained. They have been used to develop diagnostic assays by conjugating them to either quantum dots [82] or carboxyl-magnetic beads [83].…”

Section: Foot-and-mouth Diseasementioning

confidence: 99%

Use of camel single-domain antibodies for the diagnosis and treatment of zoonotic diseases

Lafaye

2018

Comparative Immunology, Microbiology and Infectious Diseases

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A B S T R A C TCamelids produce both conventional heterotetrameric antibodies and homodimeric heavy-chain only antibodies. The antigen-binding region of such homodimeric heavy-chain only antibodies consists of one single domain, called VHH. VHHs provide many advantages over conventional full-sized antibodies and currently used antibody-based fragments (Fab, scFv), including high specificity, stability and solubility, and small size, allowing them to recognize unusual antigenic sites and deeply penetrate tissues. Since their discovery, VHHs have been used extensively in diagnostics and therapy. In recent decades, the number of outbreaks of diseases transmissible from animals to humans has been on the rise. In this review, we evaluate the status of VHHs as diagnostic and therapeutic biomolecular agents for the detection and treatment of zoonotic diseases, such as bacterial, parasitic, and viral zoonosis. VHHs show great adaptability to inhibit or neutralize pathogenic agents for the creation of multifunctional VHH-based diagnostic and therapeutic molecules against zoonotic diseases.

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Characterization of single-domain antibodies against Foot and Mouth Disease Virus (FMDV) serotype O from a camelid and imaging of FMDV in baby hamster kidney-21 cells with single-domain antibody-quantum dots probes

Cited by 12 publications

References 41 publications

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus

Selection and Characterization of CSFV-Specific Single-Domain Antibodies and Their Application along with Immunomagnetic Nanobeads and Quantum Dots

Use of camel single-domain antibodies for the diagnosis and treatment of zoonotic diseases

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