Single-Domain Antibodies As Versatile Affinity Reagents for Analytical and Diagnostic Applications

González‐Sapienza, Gualberto; Rossotti, Martín A.; Rosa, Sofía Tabares-da

doi:10.3389/fimmu.2017.00977

Cited by 142 publications

(137 citation statements)

References 108 publications

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“…In addition, sdAb is easily selected and cloned by selection systems such as phage, yeast, or ribosomal display (Harmsen and De Haard 2007;Muyldermans et al 2009). These attributes make sdAbs suitable for many medical, biotechnological, diagnostic, and therapeutic applications (Arbabi-Ghahroudi 2017; Bannas et al 2017;Beghein and Gettemans 2017;Gonzalez-Sapienza et al 2017).…”

Section: Introductionmentioning

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: Introductionmentioning

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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“…Over the decades, sdAbs, more frequently referred to be as nanobodies, has been described as a credible next-generation antibody-derived biologics with significant as advances over conventional antibodies [30]. e sdAbs produced by Camelidae that has only one variable region (heavy chain), with a reduced size, and show greater stability under a range of conditions for medical and technological applications [12,31]. In contrast to McAb, lymphocytes from the PBMC pool of immunized camels should be a good source of Agbinding sdAb fragments for the construction of a large immune library.…”

Section: Discussionmentioning

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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“…The antigen binding domain of Ig-NARs or HCAbs involves a single domain only, referred to as V-NAR when derived from Ig-NARs and VHHs (nanobodies) when derived from HCAbs. The nanobodies have proven to be powerful tools in diagnostics due to their unique characteristics [135]. In particular, the recombinant expression of nanobodies in microbial systems and straightforward purification using His-tag by immobilised metal affinity chromatography makes their purification easy and very cheap.…”

Section: Prospects Of Nanobodies For Use In Immunoassaysmentioning

confidence: 99%

Paradigm shift in the diagnosis of peste des petits ruminants: scoping review

et al. 2020

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Peste des petits ruminants virus causes a highly contagious disease, which poses enormous economic losses in domestic animals and threatens the conservation of wild herbivores. Diagnosis remains a cornerstone to the Peste des petits ruminants Global Control and Eradication Strategy, an initiative of the World Organisation for Animal Health and the Food and Agriculture Organisation. The present review presents the peste des petits ruminants diagnostic landscape, including the practicality of commercially available diagnostic tools, prototype tests and opportunities for new technologies. The most common peste des petits ruminants diagnostic tools include; agar gel immunodiffusion, counter-immunoelectrophoresis, enzyme-linked immunosorbent assays, reverse transcription polymerase chain reaction either gel-based or real-time, reverse transcription loop-mediated isothermal amplification, reverse transcription recombinase polymerase amplification assays, immunochromatographic lateral flow devices, luciferase immunoprecipitation system and pseudotype-based assays. These tests vary in their technical demands, but all require a laboratory with exception of immunochromatographic lateral flow and possibly reverse transcription loop-mediated isothermal amplification and reverse transcription recombinase polymerase amplification assays. Thus, we are proposing an efficient integration of diagnostic tests for rapid and correct identification of peste des petits ruminants in endemic zones and to rapidly confirm outbreaks. Deployment of pen-side tests will improve diagnostic capacity in extremely remote settings and susceptible wildlife ecosystems, where transportation of clinical samples in the optimum cold chain is unreliable.

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Single-Domain Antibodies As Versatile Affinity Reagents for Analytical and Diagnostic Applications

Cited by 142 publications

References 108 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

Paradigm shift in the diagnosis of peste des petits ruminants: scoping review

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