A Rapid and Cost-Effective Method for the Purification of Streptococcus pneumoniae Antiserum Using Tangential Flow Filtration Method Rapid method for purification of S. Pneumoniae antiserum
Trends in Peptide and Protein Sciences,
Vol. 5 (2020),
1 Dey 2020
,
Page 1-6 (e9)
https://doi.org/10.22037/tpps.v5i.33390
Abstract
Antibodies are important agents in the laboratory diagnosis of various microorganisms such as Streptococcus pneumoniae. In this study, we prepared and purified IgG antibody against Streptococcus pneumoniae using novel methods to be used in ELISA and agglutination diagnostic kits. First, Streptococcus pneumoniae was cultured, harvested, and inactivated. Then, bacteria were injected into four mature New Zealand white rabbits, and antisera were developed. Afterward, immunoglobulins were purified using ammonium sulfate precipitation, diafilteration using Tangential Flow Filtration, and ion-exchange chromatography. The purified antibody was then biotinylated and used in ELISA. Gel electrophoresis results showed that the antibody was highly pure. Agglutination test on the primary antigen and clinical samples was four-plus. ELISA results showed that the sensitivity and specificity of the test were 95% and 100%, respectively. Results indicated that our fast method was suitable for anti-Streptococcus pneumoniae IgG purification. Repetitive qualitative and quantitative experiments confirmed high purity of the immunoglobulin. Thus, it could be a suitable candidate to be used in laboratory diagnostic kits.
HIGHLIGHTS
- A rapid and cost-effective method for the purification of Streptococcus pneumoniae antiserum.
- IgG antibody was prepared and purified by using tangential flow filtration method.
- The purified antibody can be used in ELISA and agglutination diagnostic kits.
- Agglutination
- Antigen
- ELISA
- Immunoglobin G
- Streptococcus pneumoniae
How to Cite
References
Arbique, J.C., Poyart, C., Trieu-Cuot, P., Quesne, G., Maria da Glória, S.C., Steigerwalt, A.G., Morey, R.E., Jackson, D., Davidson, R.J. and R.R. Facklam, (2004). ʺAccuracy of phenotypic and genotypic testing for identification of Streptococcus pneumoniae and description of Streptococcus pseudopneumoniae sp. nov.ʺ Journal of Clinical Microbiology, 42(10): 4686-4696.
Austrian, R. (1981). ʺSome observations on the pneumococcus and on the current status of pneumococcal disease and its prevention.ʺ Reviews of Infectious Diseases, 3(Supp. 1):S1-7.
Austrian, R. (1999). ʺThe Pneumococcus at the Millennium: Not down, not out.ʺ The Journal of Infectious Diseases, 179(Supp. 2): S338-S341.
Centers for Disease Control and Prevention (CDC) (2018). ʺGlobal Pneumococcal disease and vaccine.ʺ Retrieved from: https://www.cdc.gov/pneumococcal/global.html.
Glover, R.L. and L.J. Korschgen, (1980). ʺEvaluation of two methods of preparing antisera for precipitin tests.ʺ Wildlife Society Bulletin, 8(2): 118–122.
Hanly, W. C., Artwohl, J. E. and B. T. Bennett, (1995). ʺReview of polyclonal antibody production procedures in mammals and poultry.ʺ ILAR Journal, 37(3): 93-118.
Hendriksen, C. and J. Hau, (2002). ʺProduction of polyclonal and monoclonal antibodies.ʺ, In: Hau, J. and G. L. Van Hoosier (Eds), Handbook of Laboratory Animal Science, Second Edition: Essential Principles and Practices. CRC Press, Boca Raton, pp. 391-412.
Lalitha, M.K., Pai, R., John, T.J., Thomas, K., Jesudason, M.V., Brahmadathan, K.N., Sridharan, G. and M.C. Steinhoff, (1996). ʺSerotyping of Streptococcus pneumoniae by agglutination assays: a cost-effective technique for developing countries.ʺ Bulletin of the World Health Organization, 74(4): 387–390.
Leenaars, M. and C. F. M. Hendriksen, (2005). ʺCritical steps in the production of polyclonal and monoclonal antibodies: Evaluation and recommendations.ʺ ILAR Journal, 46(3): 269-279.
Lipman, N.S., Jackson, L.R., Trudel, L.J. and F. Weis-Garcia, (2005). ʺMonoclonal versus polyclonal antibodies: distinguishing characteristics, applications, and information resources.ʺ ILAR journal, 46(3): 258-268.
İşeri, M.N., Dinleyici, M., Kılıç, Ö., Ceyhan, M., Gürler, N. and E.Ç. Dinleyici, (2019). ʺMeningitis due to Streptococcus pneumoniae serotype 24B in a patient with cochlear implant previously vaccinated with the pneumococcal vaccine.ʺ Mikrobiyoloji Bulteni, 53(4): 451-456.
Obaro, S. and R. Adegbola, (2002). ʺThe pneumococcus: carriage, disease and conjugate vaccines.ʺ Journal of Medical Microbiology, 51(2): 98-104.
Ostler, E.L., Resmini, M., Brocklehurst, K. and G. Gallacher, (2002). ʺPolyclonal catalytic antibodies.ʺ Journal of Immunological Methods, 269(1-2): 111-124.
Parker, A.R., Lock, E., Iftikhar, A., Barber, R., Stubbs, P.D., Harding, S. and G.L. Wallis, (2017). ʺPurification and characterization of anti-pneumococcal capsular polysaccharide IgG immunoglobulins.ʺ Clinical Biochemistry, 50(1-2): 80-83.
Paton, J.C. and C. Trappetti, (2019). ʺStreptococcus pneumoniae capsular polysaccharide.ʺ In: Fischetti, V.A., Novick, R.P., Ferretti, J.J., Portnoy, D.A., Braunstein, M. and J.I. Rood (Eds), Gram‐Positive Pathogens, ASM Press, Canad, pp. 1-15.
Porter, B. D., Ortika, B. D. and C. Satzke, (2014). ʺCapsular serotyping of Streptococcus pneumoniae by latex agglutination.ʺ Journal of Visualized Experiments. 25(91):e51747.
Proom, H. (1943). ʺThe preparation of precipitating sera for the identification of animal species.ʺ The Journal of Pathology and Bacteriology, 55(4): 419-426.
Williams, B.G., Gouws, E., Boschi-Pinto, C., Bryce, J. and C. Dye, (2002). ʺEstimates of world-wide distribution of child deaths from acute respiratory infections.ʺ The Lancet Infectious Diseases, 2(1): 25-32.
Winegarner, J.H. and J. Wittkopp, (2020). ʺStreptococcus pneumoniae meningitis associated with over-the-counter sinus irrigation.ʺ Cureus, 12(5): e8258.
Wolfe, H.R. (1935). ʺThe effect of injection methods on the species specificity of serum precipitins.ʺ The Journal of Immunology, 29(1): 1-11.
World Health Organization (WHO) (2005). ʺ International travel and health: Pneumococcal disease.ʺ Retrieved from: https://www.who.int/ith/diseases/pneumococcal/en/
- Abstract Viewed: 360 times
- PDF Downloaded: 280 times