Article history: Received: 19-01-2018, Accepted: 24-05-2018, Published online: 05-07-2018
Corresponding author: Dwi Desmiyeni Putri
E-mail: firstname.lastname@example.orgCitation: Putri DD, Handharyani E, Soejoedono RD, Setiyono A, Poetri ON (2018) Production and characterization of Newcastle disease antibody as a reagent to develop a rapid immunodiagnostic test tool, Veterinary World, 11(7): 895-901.
Aim: This research was conducted to produce and characterize ND antibody as reagent candidate to develop a rapid immunodiagnostic test tool.
Materials and Methods: Four New Zealand White rabbits were used in this study and divided into two groups. First group was injected by Sato ND antigen, and second group was injected by genotype VII ND antigen. This study is divided into three steps: (a) ND antibody production, (b) ND antibody purification, and (c) ND antibody characterization. First group was rabbit injected by Sato NDV (5x108.25 egg lethal doses (ELD)50/ml) and second group was injected by genotype VII NDV (5x106.5 ELD50/ml). Antigen induction was performed by subcutaneous administrated for first (day 1) and second (day 14) injection and intravenous administrated for third (day 30) injection. Blood was collected on day 8 after third injection.
Results: Antibody production increased on second antigen injection and reached a peak on day 9 after second antigen injection. Sato and genotype VII ND antibody can be produced without adjuvant within 38 days with the highest titer 210. Based on antibody titer data, both antigens induced antibody production in a similar trend. The characterization antibody by SDS-PAGE indicated that molecular weight of immunoglobulin G (IgG) is 154.93 kDa (whole IgG), heavy chain 54.39 kDa, and light chain 27.74 kDa. ND antibodies have specificity to homologous and heterologous NDVs in varying virulence.
Conclusion: Sato and genotype VII ND antibodies have been successfully produced within 38 days without adjuvant. Specificity of ND antibodies to NDVs in varying virulence and cross-reaction between Sato ND antibody and genotype VII ND antibody indicates that the characterized ND antibodies can be used as a reagent to develop rapid immunodiagnostic test tools.
Keywords: antibody, cross reaction, reagent, SDS-PAGE.
1. Ashraf, A. and Shah, M.S. (2014) Newcastle disease: Present status and future challenges for developing countries. Afr. J. Microbiol. Res., 8: 411-416. [Crossref]
3. Al-Habeeb, M.A., Mohamed, M.H.A. and Sharawi, S. (2013) Detection and characterization of Newcastle disease virus in clinical samples using real-time RT-PCR and melting curve analysis based on matrix and fusion genes amplification. Vet. World, 6: 239-243. [Crossref]
4. Diel, D.G., Susta, L., Cardenas, G.S., Killian, M.L., Brown, C.C., Miller, P.J. and Afonso, C.L. (2012) Complete genome and clinicopathological characterization of a virulent Newcastle disease virus isolate from South America. J. Clin. Microbiol., 50: 378-387. [Crossref] [PubMed] [PMC]
5. Seal, B.S., King D.J. and Sellers H.S. (2000) The avian response to Newcastle disease virus. Dev. Com. Immunol., 24: 257-268. [Crossref]
6. Naveen, K.A., Singh, S.D., Kataria, J.M., Barathidasan, R. and Dhama, K. (2013) Detection and differentiation of pigeon paramyxovirus serotype-1(PPMV-1) isolates by RT-PCR and restriction enzyme analysis. Trop. Anim. Health. Prod., 45: 1231-1236. [Crossref] [PubMed]
7. Abdisa, T. and Tagesu, T (2013) Review on Newcastle disease of poultry and it's public health importance. J. Vet. Sci. Technol., 8(3): 1-7.
8. Cattoli, G., Susta, L., Terregino, C. and Brown, C. (2011) Newcastle disease: A review of field recognition and current methods of laboratory detection. J. Vet. Diag. Invest., 23(4): 637-656. [Crossref] [PubMed]
9. Office International des Epizooties (OIE). (2012) Newcastle Disease. OIE Terestrial Manual, France. p1-19.
10. Choi, K.S., Kye, S.J., Kim, J.Y., To, T.L., Nguyen, D.T., Lee, Y.J. and Lee, H.S. (2014) Molecular epidemiology of Newcastle disease viruses in Vietnam. Trop. Anim. Health Prod., 46(1): 271-277. [Crossref] [PubMed]
11. Putri, D.D., Handharyani, E., Soejono, R.D., Setiyono, A., Mayasari, N.I. and Poetri, O.N. (2017) Pathotypic characterization of Newcastle disease virus isolated from vaccinated chicken in West Java Indonesia. Vet. World, 10(4): 438-444. [Crossref] [PubMed] [PMC]
12. Putri, D.D., Handharyani, E., Soejono, R.D., Setiyono, A. and Poetri, O.N. (2018) Genotypic characterization of Newcastle disease virus isolated from commercial flock chicken in West Java Indonesia. Pak. Vet. J., 38(2): 184-188.
14. Johnstone, A. and Thorpe, R. (1988) Immunochemistry in Practice. Blackwell Scienti?c Publications, Oxford, England.
15. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. (2003) Current Protocols in Molecular Biology. John Willey & Sons, New York, US.
16. Sambrook, J. and Russel, D.W. (2001) Molecular Cloning: A Laboratory Manual. Vol. 1. Cold Spring Harbour Lab Press, New York, USA.
17. Bergmann-Leitner, E.S., Mease, R.M., Duncan, E.H., Khan, F., Waitumbi, J. and Angov, E. (2008) Evaluation of immunoglobulin purification methods and their impact on quality and yield of antigen-specific antibodies. Malar J., 7: 129. [Crossref] [PubMed] [PMC]
18. Green, M.R. and Sambrook, J. (2012) Molecular Cloning - A Laboratory Manual. 4th ed. Cold Spring Habor Laboratory Press, Cold Spring Harbor, NY.
19. Nandin, I.S., Fong, C., Deantonio, C., Torreno-Pina, J.A., Pecetta, S., Maldonado, P., Gasparrini, F., Ordovas-Montanes, J., Kazer, S.W., Kjaer, S., Borley, D.W., Nair, U., Coleman, J.A., Lingwood, D., Shalek, A.K., Meffre, E., Poignard, P., Burton, D.R. and Batista, F.D. (2017) Novel in vitro booster vaccination to rapidly generate antigen-specific human monoclonal antibodies. J. Exp. Med., 2017: 1-20. [Crossref]
21. Wibawan, I.W.T. and Soejoedono, R.D. (2013) Intisari Imunologi Medis. Faculty of Veterinary Medicine Press,Bogor, Indonesia. p35.
23. Hanley, W.C., Artwohl, J. and Bennett, B.T. (1995) Review of polyclonal antibody production procedures in mammals and poultry. Inst. Lab. Anim. Res. J., 37(3): 91-118. [Crossref]
25. Howard, G.C. and Kaser, M.R. (2006) Making and Using Antibodies: A Practical Handbook. CRC Press, Boca Raton. [Crossref]
27. Samiullah, M., Rizvi, F., Anjun, A.D. and Shah, M.F.A. (2006) Rising hyperimmune serum against avian paramyxovirus (APMV-1) and pigeon paramyxovirus (PPMV-1) in rabbits and their cross-reactivity. Pak. J. Biol. Sci., 9: 2184-2186. [Crossref]
28. Murphy, C., Devine, T. and O'Kennedy, R. (2016) Technology advancements in antibody purification. Dovepress, 2016: 17-32. Available from: https://www.com/technology-advancements-in-antibody-purification-peer-reviewed-full. Accessed on 21-07-2017.
29. Wingfield, P.T. (2001) Protein precipitation using ammonium sulfate. Curr. Protoc. Protein Sci. Appendix-3F. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817497. Accessed on 05-04-2017.
30. Page, M. and Thorppe, R. (2001) Purification of IgG by Precipitation with Sodium Sulfate or Ammonium Sulfate. In: Hage, D.S. editor. Handbook of Affinity Chromatography Book Chapter. 2nd ed. Taylor and Francis Group. New York. Available from: https://www.books.google.co.id/books?id. Accessed on 05-07-2016.
31. Jan-Roblero, J., Garcia-Gomez, E., Rodriquez-Martinez, S., Cancino-Diaz, M.E. and Cancino-Diaz, J.C. (2017) Surface Proteins of Staphylococcus aureus. Chapter of book The Rise of Virulence and Antibiotic Resistance in Staphylococcus aureus. Intech. Avaialble from: https://www.cdn.intechopen.com/pdfs-wm/53157. Accessed on 15-11-2017. [Crossref]
32. Ghose, S., Allen, M., Hubbard, B., Brooks, C. and Cramer, S.M. (2005) Antibody variable region interactions with Protein A: Implications for the development of generic purification processes. Biotechnol. Bioeng., 92(6): 665-673. [Crossref] [PubMed]
33. Johnson, M. (2013) Rabbit Antibody. Available from: https://www.labome.com/method/Rabbit-Antibody.html. Accessed on 23-01-2017. [Crossref]
34. De-Buysscher, E.V. and Patterson, R.M. (1995) In: Burgess, G.W., editor. Immune Respons. Gadjah Mada University Press, Yogyakarta. p1-9.
35. Tizard, I.R. (2004) An Introduction to Veterinary Immunology. 7th ed. Elsevier, Philadelphia.
36. Yang, L., Biswas, M.E. and Chen, P. (2003) Study of binding between protein a and immunoglobulin g using a surface tension probe. Biophys. J., 84: 509-522. [Crossref]
37. Zhang, W.D., Wang, W.H. and Jia, S. (2015) Distribution of immunoglobulin G antibody secretory cells in small intestine of Bactrian camels (Camelus bactrianus). Vet. Res., 11: 222. [Crossref]38. Frank, S.A. (2002) Immunology and Evolution of Infectious Disease. Princeton University Press, Princeton, NJ.