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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.

Wang D, Yang S, Yin S, Shang Y, Du P, Guo J, He J, Cai J, Liu X - BMC Vet. Res. (2015)

Bottom Line: Results demonstrated that the two sdAbs were specific for serotype O and shared no cross-reactivity with serotypes A and Asia 1.The equilibrium dissociation constant (KD) values of the two sdAbs ranged from 6.23 nM to 8.24 nM, which indicated high affinity to FMDV antigens.Co-localization with the sdAb-AF488 and sdAb-QD probes indicated the same location of FMDV virions in baby hamster kidney-21 (BHK-21) cells. sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China. sdau_dw@163.com.

ABSTRACT

Background: Foot-and-mouth disease (FMD) is a highly contagious disease that affects cloven-hoofed animals and causes significant economic losses to husbandry worldwide. The variable domain of heavy-chain antibodies (VHHs or single domain antibodies, sdAbs) are single-domain antigen-binding fragments derived from camelid heavy-chain antibodies.

Results: In this work, two sdAbs against FMD virus (FMDV) serotype O were selected from a camelid phage display immune library and expressed in Escherichia coli. The serotype specificity and affinity of the sdAbs were identified through enzyme-linked immunosorbent assay and surface plasmon resonance assay. Moreover, the sdAbs were conjugated with quantum dots to constitute probes for imaging FMD virions. Results demonstrated that the two sdAbs were specific for serotype O and shared no cross-reactivity with serotypes A and Asia 1. The equilibrium dissociation constant (KD) values of the two sdAbs ranged from 6.23 nM to 8.24 nM, which indicated high affinity to FMDV antigens. Co-localization with the sdAb-AF488 and sdAb-QD probes indicated the same location of FMDV virions in baby hamster kidney-21 (BHK-21) cells.

Conclusions: sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.

No MeSH data available.


Related in: MedlinePlus

Serotype specificity identification of sdAbs against FMDV serotype O, Asia 1, and A antigens. All samples were tested for absorbance at 450 nm. The negative camel serum was coated as the negative control, and the polyclonal antibodies against FMDV type O were coated as the positive control. Error bars represent the ± standard deviation of three independent measurements
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Fig6: Serotype specificity identification of sdAbs against FMDV serotype O, Asia 1, and A antigens. All samples were tested for absorbance at 450 nm. The negative camel serum was coated as the negative control, and the polyclonal antibodies against FMDV type O were coated as the positive control. Error bars represent the ± standard deviation of three independent measurements

Mentions: The results of sandwich ELISA showed that the two sdAbs displayed high affinities to FMDV type O and exhibited no cross-reactivity with serotypes Asia 1 and A in ELISA (Fig. 6). The specificities and high affinities of sdAbs to FMDV serotype O indicated that they could be effective antibodies to this target FMDV serotype.Fig. 6


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 D, Yang S, Yin S, Shang Y, Du P, Guo J, He J, Cai J, Liu X - BMC Vet. Res. (2015)

Serotype specificity identification of sdAbs against FMDV serotype O, Asia 1, and A antigens. All samples were tested for absorbance at 450 nm. The negative camel serum was coated as the negative control, and the polyclonal antibodies against FMDV type O were coated as the positive control. Error bars represent the ± standard deviation of three independent measurements
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4489003&req=5

Fig6: Serotype specificity identification of sdAbs against FMDV serotype O, Asia 1, and A antigens. All samples were tested for absorbance at 450 nm. The negative camel serum was coated as the negative control, and the polyclonal antibodies against FMDV type O were coated as the positive control. Error bars represent the ± standard deviation of three independent measurements
Mentions: The results of sandwich ELISA showed that the two sdAbs displayed high affinities to FMDV type O and exhibited no cross-reactivity with serotypes Asia 1 and A in ELISA (Fig. 6). The specificities and high affinities of sdAbs to FMDV serotype O indicated that they could be effective antibodies to this target FMDV serotype.Fig. 6

Bottom Line: Results demonstrated that the two sdAbs were specific for serotype O and shared no cross-reactivity with serotypes A and Asia 1.The equilibrium dissociation constant (KD) values of the two sdAbs ranged from 6.23 nM to 8.24 nM, which indicated high affinity to FMDV antigens.Co-localization with the sdAb-AF488 and sdAb-QD probes indicated the same location of FMDV virions in baby hamster kidney-21 (BHK-21) cells. sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China. sdau_dw@163.com.

ABSTRACT

Background: Foot-and-mouth disease (FMD) is a highly contagious disease that affects cloven-hoofed animals and causes significant economic losses to husbandry worldwide. The variable domain of heavy-chain antibodies (VHHs or single domain antibodies, sdAbs) are single-domain antigen-binding fragments derived from camelid heavy-chain antibodies.

Results: In this work, two sdAbs against FMD virus (FMDV) serotype O were selected from a camelid phage display immune library and expressed in Escherichia coli. The serotype specificity and affinity of the sdAbs were identified through enzyme-linked immunosorbent assay and surface plasmon resonance assay. Moreover, the sdAbs were conjugated with quantum dots to constitute probes for imaging FMD virions. Results demonstrated that the two sdAbs were specific for serotype O and shared no cross-reactivity with serotypes A and Asia 1. The equilibrium dissociation constant (KD) values of the two sdAbs ranged from 6.23 nM to 8.24 nM, which indicated high affinity to FMDV antigens. Co-localization with the sdAb-AF488 and sdAb-QD probes indicated the same location of FMDV virions in baby hamster kidney-21 (BHK-21) cells.

Conclusions: sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.

No MeSH data available.


Related in: MedlinePlus