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

BMC Veterinary Research

Xiangtao Liu¹  Jianping Cai¹  Jijun He¹  Jianhong Guo¹  Ping Du¹  Youjun Shang¹  Shuanghui Yin¹  Shunli Yang¹  Di Wang¹ 

[1] 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

关键词: Imaging;    Probe;    Quantum dots;    Single domain antibody;    Immune library;    Foot-and-mouth disease virus serotype O;   

Others  :  1206108
DOI  :  10.1186/s12917-015-0437-2

 received in 2014-10-13, accepted in 2015-05-12,  发布年份 2015

【 摘 要 】

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.

【 授权许可】

   
2015 Wang et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Yang PC, Chu RM, Chung WB, Sung HT. Epidemiological characteristics and financial costs of the 1997 foot-and-mouth disease epidemic in Taiwan. Vet Rec. 1999; 145:731-4.
• [2]Thompson D, Muriel P, Russell D, Osborne P, Bromley A, Rowland M et al.. Economic costs of the foot and mouth disease outbreak in the United Kingdom in 2001. Rev Sci Tech. 2002; 21:675-87.
• [3]Garland A, Donaldson A. Foot-and-mouth disease. Surveillance. 1990; 17:6-8.
• [4]Bachrach HL. Foot-and-mouth disease. Ann Rev Microbiol. 1968; 22:201-44.
• [5]Hamers-Casterman C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa EB et al.. Naturally occurring antibodies devoid of light chains. Nature. 1993; 363:446-8.
• [6]Greenberg AS, Avila D, Hughes M, Hughes A, McKinney EC, Flajnik MF. A new antigen receptor gene family that undergoes rearrangement and extensive somatic diversification in sharks. Nature. 1995; 374:168-73.
• [7]Harmsen MM, De Haard HJ. Properties, production, and applications of camelid single-domain antibody fragments. Appl Microbiol Biotechnol. 2007; 77:13-22.
• [8]Rahbarizadeh F, Rasaee MJ, Forouzandeh-Moghadam M, Allameh AA. High expression and purification of the recombinant camelid anti-MUC1 single domain antibodies in Escherichia coli. Protein Expr Purif. 2005; 44:32-8.
• [9]Frenken LG, van der Linden RH, Hermans PW, Bos JW, Ruuls RC, de Geus B et al.. Isolation of antigen specific llama VHH antibody fragments and their high level secretion by Saccharomyces cerevisiae. J Biotechnol. 2000; 78:11-21.
• [10]Ismaili A, Jalali-Javaran M, Rasaee MJ, Rahbarizadeh F, Forouzandeh-Moghadam M, Memari HR. Production and characterization of anti-(mucin MUC1) single-domain antibody in tobacco (Nicotiana tabacum cultivar Xanthi). Biotechnol Appl Biochem. 2007; 47:11-9.
• [11]Revets H, De Baetselier P, Muyldermans S. Nanobodies as novel agents for cancer therapy. Expert Opinion Biol Ther. 2005; 5:111-24.
• [12]Cortez-Retamozo V, Lauwereys M, Hassanzadeh Gh G, Gobert M, Conrath K, Muyldermans S et al.. Efficient tumor targeting by single-domain antibody fragments of camels. Int J Cancer. 2002; 98:456-62.
• [13]Leutwyler WK, Bürgi SL, Burgl H. Semiconductor clusters, nanocrystals, and quantum dots. Science. 1996; 271:933-7.
• [14]Voura EB, Jaiswal JK, Mattoussi H, Simon SM. Tracking metastatic tumor cell extravasation with quantum dot nanocrystals and fluorescence emission-scanning microscopy. Nat Med. 2004; 10:993-8.
• [15]Lidke DS, Nagy P, Heintzmann R, Arndt-Jovin DJ, Post JN, Grecco HE et al.. Quantum dot ligands provide new insights into erbB/HER receptor–mediated signal transduction. Nature Biotechnol. 2004; 22:198-203.
• [16]Jaiswal JK, Mattoussi H, Mauro JM, Simon SM. Long-term multiple color imaging of live cells using quantum dot bioconjugates. Nature Biotechnol. 2002; 21:47-51.
• [17]Zaman MB, Baral TN, Zhang J, Whitfield D, Yu K. Single-domain antibody functionalized CdSe/ZnS quantum dots for cellular imaging of cancer cells. J Physical Chem C. 2008; 113:496-9.
• [18]Yin S, Yang S, Shang Y, Sun S, Zhou G, Jin Y et al.. Characterization of Asia 1 sdAb from Camels Bactrianus (C. bactrianus) and Conjugation with Quantum Dots for Imaging FMDV in BHK-21 Cells. PloS One. 2013; 8: Article ID e63500
• [19]English D, Andersen BR. Single-step separation of red blood cells. Granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J Immunol Methods. 1974; 5:249-52.
• [20]Hoogenboom HR, Griffiths AD, Johnson KS, Chiswell DJ, Hudson P, Winter G. Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucleic Acids Res. 1991; 19:4133-7.
• [21]Hayhurst A, Happe S, Mabry R, Koch Z, Iverson BL, Georgiou G. Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. J Immunol Methods. 2003; 276:185-96.
• [22]Goffinet M, Chinestra P, Lajoie-Mazenc I, Medale-Giamarchi C, Favre G, Faye JC. Identification of a GTP-bound Rho specific scFv molecular sensor by phage display selection. BMC Biotechnol. 2008; 8:34. BioMed Central Full Text
• [23]Hoogenboom HR, de Bruine AP, Hufton SE, Hoet RM, Arends JW, Roovers RC. Antibody phage display technology and its applications. Immunotechnology. 1998; 4:1-20.
• [24]Yang S, Shang Y, Yin S, Tian H, Chen Y, Sun S et al.. Selection and identification of single-domain antibody fragment against capsid protein of porcine circovirus type 2 (PCV2) from C. bactrianus. Vet Immunol Immunopathol. 2014; 160:12-9.
• [25]Yin S, Sun S, Yang S, Shang Y, Cai X, Liu X. Self-assembly of virus-like particles of porcine circovirus type 2 capsid protein expressed from Escherichia coli. Virol J. 2010; 7:166. BioMed Central Full Text
• [26]Butt TR, Edavettal SC, Hall JP, Mattern MR. SUMO fusion technology for difficult-to-express proteins. Protein Expr Purif. 2005; 43:1-9.
• [27]Liu JL, Hu ZQ, Xing S, Xue S, Li HP, Zhang JB et al.. Attainment of 15-fold higher affinity of a Fusarium-specific single-chain antibody by directed molecular evolution coupled to phage display. Mol Biotechnol. 2012; 52:111-22.
• [28]Vu KB, Ghahroudi MA, Wyns L, Muyldermans S. Comparison of llama VH sequences from conventional and heavy chain antibodies. Mol Immunol. 1997; 34:1121-31.
• [29]Zheng H, He J, Guo J, Jin Y, Yang F, Lv L et al.. Genetic characterization of a new pandemic Southeast Asia topotype strain of serotype O foot-and-mouth disease virus isolated in China during 2010. Virus Genes. 2012; 44:80-8.
• [30]Valdazo-Gonzalez B, Timina A, Scherbakov A, Abdul-Hamid NF, Knowles NJ, King DP. Multiple introductions of serotype O foot-and-mouth disease viruses into East Asia in 2010–2011. Vet Res. 2013; 44:76. BioMed Central Full Text
• [31]Bignami A, Eng LF, Dahl D, Uyeda CT. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 1972; 43:429-35.
• [32]Henle G, Henle W. Immunofluorescence in cells derived from Burkitt's lymphoma. J Bacteriol. 1966; 91:1248-56.
• [33]Rahbarizadeh F, Rasaee MJ, Forouzandeh M, Allameh AA. Over expression of anti-MUC1 single-domain antibody fragments in the yeast Pichia pastoris. Mol Immunol. 2006; 43:426-35.
• [34]Olichon A, Surrey T. Selection of genetically encoded fluorescent single domain antibodies engineered for efficient expression in Escherichia coli. J Biol Chem. 2007; 282:36314-20.
• [35]Rothbauer U, Zolghadr K, Tillib S, Nowak D, Schermelleh L, Gahl A et al.. Targeting and tracing antigens in live cells with fluorescent nanobodies. Nat Methods. 2006; 3:887-9.
• [36]Gueorguieva D, Li S, Walsh N, Mukerji A, Tanha J, Pandey S. Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis. FASEB J. 2006; 20:2636-8.
• [37]Wesolowski J, Alzogaray V, Reyelt J, Unger M, Juarez K, Urrutia M et al.. Single domain antibodies: promising experimental and therapeutic tools in infection and immunity. Med Microbiol Immunol. 2009; 198:157-74.
• [38]Pini A, Bracci L. Phage display of antibody fragments. Curr Protein Pept Sci. 2000; 1:155-69.
• [39]Hoogenboom HR. Selecting and screening recombinant antibody libraries. Nat Biotechnol. 2005; 23:1105-16.
• [40]Alexandersen S, Wernery U, Nagy P, Frederiksen T, Normann P. Dromedaries (Camelus dromedarius) are of low susceptibility to inoculation with foot-and-mouth disease virus serotype O. J Comp Pathol. 2008; 139:187-93.
• [41]Huo Q. A perspective on bioconjugated nanoparticles and quantum dots. Colloids Surf B Biointerfaces. 2007; 59:1-10.
• [42]Monaghan P, Cook H, Jackson T, Ryan M, Wileman T. The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells. J Gen Virol. 2004; 85:933-46.