Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus
Abstract 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 loss...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Yang, Shunli [verfasserIn] |
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| Format: |
Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2018 |
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| Schlagwörter: |
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| Anmerkung: |
© Springer-Verlag GmbH Germany, part of Springer Nature 2018 |
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| Übergeordnetes Werk: |
Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 102(2018), 20 vom: 24. Aug., Seite 8931-8942 |
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| Übergeordnetes Werk: |
volume:102 ; year:2018 ; number:20 ; day:24 ; month:08 ; pages:8931-8942 |
| Links: |
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| DOI / URN: |
10.1007/s00253-018-9324-7 |
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OLC2050796064 |
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| 520 | |a Abstract 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. | ||
| 650 | 4 | |a Single domain antibody | |
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| 650 | 4 | |a Immunomagnetic nanobeads | |
| 650 | 4 | |a Porcine epidemic diarrhea virus | |
| 700 | 1 | |a Li, Li |4 aut | |
| 700 | 1 | |a Yin, Shuanghui |4 aut | |
| 700 | 1 | |a Shang, Youjun |4 aut | |
| 700 | 1 | |a Khan, Muhammad Umar Zafar |4 aut | |
| 700 | 1 | |a He, Xueyang |4 aut | |
| 700 | 1 | |a Yuan, Li |4 aut | |
| 700 | 1 | |a Gao, Xue |4 aut | |
| 700 | 1 | |a Liu, Xiangtao |4 aut | |
| 700 | 1 | |a Cai, Jianping |4 aut | |
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10.1007/s00253-018-9324-7 doi (DE-627)OLC2050796064 (DE-He213)s00253-018-9324-7-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Yang, Shunli verfasserin aut Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus 2018 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract 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. Single domain antibody Quantum dots probe Immunomagnetic nanobeads Porcine epidemic diarrhea virus Li, Li aut Yin, Shuanghui aut Shang, Youjun aut Khan, Muhammad Umar Zafar aut He, Xueyang aut Yuan, Li aut Gao, Xue aut Liu, Xiangtao aut Cai, Jianping aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 102(2018), 20 vom: 24. Aug., Seite 8931-8942 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 https://doi.org/10.1007/s00253-018-9324-7 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 102 2018 20 24 08 8931-8942 |
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10.1007/s00253-018-9324-7 doi (DE-627)OLC2050796064 (DE-He213)s00253-018-9324-7-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Yang, Shunli verfasserin aut Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus 2018 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract 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. Single domain antibody Quantum dots probe Immunomagnetic nanobeads Porcine epidemic diarrhea virus Li, Li aut Yin, Shuanghui aut Shang, Youjun aut Khan, Muhammad Umar Zafar aut He, Xueyang aut Yuan, Li aut Gao, Xue aut Liu, Xiangtao aut Cai, Jianping aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 102(2018), 20 vom: 24. Aug., Seite 8931-8942 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 https://doi.org/10.1007/s00253-018-9324-7 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 102 2018 20 24 08 8931-8942 |
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10.1007/s00253-018-9324-7 doi (DE-627)OLC2050796064 (DE-He213)s00253-018-9324-7-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Yang, Shunli verfasserin aut Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus 2018 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract 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. Single domain antibody Quantum dots probe Immunomagnetic nanobeads Porcine epidemic diarrhea virus Li, Li aut Yin, Shuanghui aut Shang, Youjun aut Khan, Muhammad Umar Zafar aut He, Xueyang aut Yuan, Li aut Gao, Xue aut Liu, Xiangtao aut Cai, Jianping aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 102(2018), 20 vom: 24. Aug., Seite 8931-8942 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 https://doi.org/10.1007/s00253-018-9324-7 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 102 2018 20 24 08 8931-8942 |
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10.1007/s00253-018-9324-7 doi (DE-627)OLC2050796064 (DE-He213)s00253-018-9324-7-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Yang, Shunli verfasserin aut Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus 2018 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract 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. Single domain antibody Quantum dots probe Immunomagnetic nanobeads Porcine epidemic diarrhea virus Li, Li aut Yin, Shuanghui aut Shang, Youjun aut Khan, Muhammad Umar Zafar aut He, Xueyang aut Yuan, Li aut Gao, Xue aut Liu, Xiangtao aut Cai, Jianping aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 102(2018), 20 vom: 24. Aug., Seite 8931-8942 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 https://doi.org/10.1007/s00253-018-9324-7 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 102 2018 20 24 08 8931-8942 |
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10.1007/s00253-018-9324-7 doi (DE-627)OLC2050796064 (DE-He213)s00253-018-9324-7-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Yang, Shunli verfasserin aut Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus 2018 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature 2018 Abstract 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. Single domain antibody Quantum dots probe Immunomagnetic nanobeads Porcine epidemic diarrhea virus Li, Li aut Yin, Shuanghui aut Shang, Youjun aut Khan, Muhammad Umar Zafar aut He, Xueyang aut Yuan, Li aut Gao, Xue aut Liu, Xiangtao aut Cai, Jianping aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 102(2018), 20 vom: 24. Aug., Seite 8931-8942 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 https://doi.org/10.1007/s00253-018-9324-7 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 102 2018 20 24 08 8931-8942 |
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Enthalten in Applied microbiology and biotechnology 102(2018), 20 vom: 24. Aug., Seite 8931-8942 volume:102 year:2018 number:20 day:24 month:08 pages:8931-8942 |
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single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus |
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Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus |
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Abstract 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. © Springer-Verlag GmbH Germany, part of Springer Nature 2018 |
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Abstract 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. © Springer-Verlag GmbH Germany, part of Springer Nature 2018 |
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Abstract 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. © Springer-Verlag GmbH Germany, part of Springer Nature 2018 |
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