Bacterial Lipid Modification of ICP11 and a New ELISA System Applicable for WSSV Infection Detection

Original Article
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Abstract

In ELISA, a popular analytical diagnostic tool, the stable non-covalent immobilization (coating) of hydrophilic proteins/peptides on to hydrophobic polystyrene surface has remained a major common challenge. Recombinant bacterial lipid modification of proteins in Escherichia coli system has been shown in this study to solve this problem owing to the hydrophobic anchorage provided by three fatty acyl groups in N-acyl-S-diacylglyceryl Cys at the N-terminus. Exploiting this first post-translational protein engineering, the most abundantly expressed white spot syndrome viral protein ICP11 was lipid-modified and tested as a new target in a new ELISA method useful to shrimp farming. The lipid served as a potent adjuvant to enhance the titer (16 times) of higher affinity antibodies where amino terminal lipoamino acid N-acyl-S-diacylglyceryl cysteine of bacterial lipoproteins induce inflammatory responses through TLR and stimulate humoral immune responses without additional adjuvant and also aided in the immobilization of even a few nanograms of ICP11. Competition between the immobilized and the free antigen from the sample provided a sensitive measure of antigen in the infected shrimp tissues. The detection limit for ICP11 protein using competitive ELISA was 250 pg and the linear range of the assay was 15–240 ng.

Keywords

Lipid modification Shrimp WSSV ICP11 ELISA 

Notes

Acknowledgements

The authors thank Dr. S. Kumar for providing the polyclonal antiserum and Dr. Shrikrishnan Sankaran for affinity quantification and valuable comments. The authors dedicate this article to (late) Dr. V. Murugan, Centre for Biotechnology, Anna University, Chennai.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.

References

  1. Ayub F, Md YS, Md SA (2008) Prevalance of white spot syndrome virus infection detected by one-step and nested PCR in selected tiger shrimp (Penaeus monodon) hatcheries. Aquac Int 16:405–415CrossRefGoogle Scholar
  2. Chaivisuthangkura P, Longyant S, Rukpratanporn S, Srisuk C, Sridulyakul P, Sithigorngul P (2010) Enhanced white spot syndrome virus (WSSV) detection sensitivity using monoclonal antibody specific to heterologously expressed VP 19 envelope protein. Aquaculture 299:15–20CrossRefGoogle Scholar
  3. Chang PS, Lo CC, Wang YC, Kou GH (1996) Identification of white spot syndrome associated baculovirus (WSBV) target organs in the shrimp Penaeus monodon by in situ hybridization. Dis Aquat Org 27:131–139CrossRefGoogle Scholar
  4. Cuéllar-Anjel J, White-Noble B, Schofield P, Chamorro R, Lightner DV (2012) Report of significant WSSV-resistance in the Pacific white shrimp, Litopenaeus vannamei, from a Panamanian breeding program. Aquaculture 368–369:36–39CrossRefGoogle Scholar
  5. Durand SV, Redman RM, Mohney LL, Tang-Nelson K, BOnami JR, Lightner DV (2003) Qualitative and quantitative studies on the relative virus load of tails and heads of shrimp acutely infected with WSSV. Aquaculture 216:9–18CrossRefGoogle Scholar
  6. Kamalakannan S, Murugan V, Jagannadham MV, Nagaraj R, Sankaran K (2004) Bacterial lipid modification of proteins for novel protein engineering applications. Protein Eng Des Sel 17:721–729CrossRefGoogle Scholar
  7. Kumar S, Balamurali MM, Sankaran K (2014) Bacterial lipid modification of proteins requires appropriate secretory signals even for expression—implications for biogenesis and protein engineering. Mol Membr Biol 31(6):183–194CrossRefPubMedGoogle Scholar
  8. Leal CAG, Carvalho-Castro GA, Cottorello AC, Leite RC, Figueiredo HCP (2013) Comparative analysis of conventional PCR and real-time PCR to diagnose shrimp WSD. Braz J Microbiol 44(3):901–904Google Scholar
  9. Liu W, Wang YT, Tian DS, Yin ZC, Kwang J (2002) Detection of white spot syndrome virus (WSSV) of shrimp by means of monoclonal antibodies (MAbs) specific to an envelope protein (28 kDa). Dis Aquat Org 49:11–18CrossRefPubMedGoogle Scholar
  10. Lo CF, Ho CH, Peng SE, Chen CH, Hsu HC, Chiu YL, Chang CF, Lui KF, Su MS, Wang CH, Kou GH (1996) White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp, crabs and other arthropods. Dis Aquat Org 27:215–225CrossRefGoogle Scholar
  11. Mendoza-Cano F, Sanchez-Paz A (2013) Development and validation of a quantitative real-time polymerase chain assay for universal detection of the white spot syndrome virus in marine crustaceans. Virol J 10(186):1–11Google Scholar
  12. Nagalakshmi G, Jyothi S (2013) White spot syndrome virus detection in shrimp images using image segmentation techniques. Int J Adv Res Comput Sci Softw Eng 3:107–112Google Scholar
  13. Patil R, Palaksha KJ, Anil TM, Guruchannabasavanna PP, Shankar KM, Mohan CV, Sripada A (2008) Evaluation of an immunodot test to manage white spot syndrome virus (WSSV) during cultivation of the giant tiger shrimp Penaeus monodon. Dis Aquat Org 79:157–161CrossRefPubMedGoogle Scholar
  14. Reddy AD, Jeyasekaran G, Shakila RJ (2013) Morphogenesis, pathogenesis, detection and transmission risks of white spot syndrome virus in shrimps. Fish Aquac J 66:1–10Google Scholar
  15. Rout N, Citarasu T, Ravindran R, Murugan V (2005) Transcriptional and translational expression profile of a white spot syndrome viral (WSSV) gene in different organs of infected shrimp. Aquaculture 245:31–38CrossRefGoogle Scholar
  16. Sharmila S, Christinia I, Kiran P, Reddy MVR, Sankaran K, Kaliraj P (2013) Bacterial lipid modification enhances immunoprophylaxis of filarial abundant larval transcript-2 protein in Mastomys model. Parasite Immunol 35:201–213CrossRefPubMedGoogle Scholar
  17. Shih HH (2002) Detection and titration of white spot syndrome virus using a Blue-cell ELISA. J Fish Dis 25:185–189CrossRefGoogle Scholar
  18. Sirwattanarat R, Longyant S, Chaivisuthangkura P, Wangman P, Vaniksampanna A, Sithigorngul P (2013) Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11. Arch Virol 158:967–979CrossRefGoogle Scholar
  19. Sithigorngul W, Rukpratanporn S, Pecharaburanin N, Lonjyant S, Chaibisuthangkura P, Sithigorngul P (2006) A simple and rapid Immunochromatographic test strip of detection of white spot syndrome virus (WSSV) of shrimp. Dis Aquat Org 72:101–106CrossRefPubMedGoogle Scholar
  20. Takahashi Y, Itami T, Maeda M, Suzuki N, Kasornchandra J, Supamattaya K, Khongpradit R, Boonyaratpalin S, Kondo M, Kawai K, Kusuda R, Hirono I, Aoki T (1996) Polymerase chain reaction (PCR) amplification of bacilliform virus (RV-PJ) DNA in Penaeus japonicus Bate and systemic ectodermal and mesodermal baculovirus (SEMBV) DNA in Penaeus monodon Fabricius. J Fish Diseases 19:399–403CrossRefGoogle Scholar
  21. Tang KFJ, Lightner DV (2000) Quantification of white spot syndrome virus DNA through a competitive polymerase chain reaction. Aquaculture 189:11–21CrossRefGoogle Scholar
  22. Thiruppathiraja C, Kumar S, Murugan V, Adaikkappan P, Sankaran K, Alagar M (2011) An enhanced immuno-dot blot assay for the detection of white spot syndrome virus in shrimp using antibody conjugated gold nanoparticles probe. Aquaculture 318:262–267CrossRefGoogle Scholar
  23. Vaseeharan B, Jayakumar R, Ramasamy P (2003) PCR-based detection of white spot syndrome virus in cultured and captured crustaceans in India. Lett Appl Microbiol 37:443–447CrossRefPubMedGoogle Scholar
  24. Wang X, Zhan W (2006) Development of an immunochromatographic test to detect white spot syndrome virus of shrimp. Aquaculture 225:196–200CrossRefGoogle Scholar
  25. Wang X, Zhan W, Xing J (2006) Development of dot-immunogold filtration assay to detect white spot syndrome virus of shrimp. J Virol Methods 132:212–215CrossRefPubMedGoogle Scholar
  26. Wang HC, Wang HC, Ko TP, Lee YM, Leu JH, Ho CH, Huang WP, Lo CF, Wang AHJ (2008) White spot syndrome virus protein ICP11: a histone-binding DNA mimic that disrupts nucleosome assembly. PNAS 105:20758–20763CrossRefPubMedPubMedCentralGoogle Scholar
  27. Xia X, Yu Y, Weidmann M, Pan Y, Yan S, Wang Y (2014) Rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay. PLoS One 9(8):1–8Google Scholar
  28. Zhan W, Wang X, Chen J, Xing J, Fukuda H (2004) Elimination of shrimp endogenous alkaline phosphatase background and development of enzyme immunoassays for the detection of white spot syndrome virus (WSSV). Aquaculture 239:15–21CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Centre for BiotechnologyAnna UniversityChennaiIndia

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