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Highly Sensitive Chemiluminescent Analysis of Residual Bovine Serum Albumin (BSA) Based on a Pair of Specific Monoclonal Antibodies and Peroxyoxalate–glyoxaline–PHPPA Dimer Chemiluminescent System in Vaccines

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Abstract

Enzyme-linked immunosorbent assay (ELISA), horseradish peroxidase (HRP)-catalyzed fluorescent reaction, and oxalate chemiluminescence analysis have been combined to develop a highly sensitive, simple, and rapid method for analysis of bovine serum albumin (BSA) based on a pair of specific monoclonal antibodies in vaccines. A typical “sandwich type” immunoassay was used. Reaction of 3-(4-hydroxyphenyl propionate) (PHPPA) with hydrogen peroxide-urea, catalyzed by HRP, produced fluorescence of 3-(4-hydroxyphenyl propionate) dimer, which was detected by chemiluminescence analysis with the bis(2,4,6-trichlorophenyl)oxalate (TCPO)–H2O2–glyoxaline–PHPPA dimer chemiluminescent system. This method exhibited high performance with a linear correlation between response and amount of bovine serum albumin (BSA) in the range 0.1 to 100.0 ng mL−1 (r = 0.9988), and the detection limit was 0.03 ng mL−1 (S/N = 3). Intra- and interassay coefficient variations were all lower than 9.0% at three concentrations (1.0, 20.0, and 80.0 ng mL−1). The proposed method has been used for successful analysis of the amount of residual BSA in vaccines. The results obtained compared well with those obtained by conventional colorimetric ELISA and luminol chemiluminescent ELISA.

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References

  1. Boye, J. I., Alli, I., & Ismail, A. A. (1996). Interactions involved in the gelation of bovine serum albumin. Journal of Agricultural and Food Chemistry, 44, 996–1004.

    Article  CAS  Google Scholar 

  2. Ferry, J. D. (1948). Protein gels. Advances in Protein Chemistry, 4, 1–78.

    Article  CAS  Google Scholar 

  3. Kratz, F., Fichtner, I., Beyer, U., Schumacher, P., Roth, T., Fiebig, H. H., & Unger, C. (1997). Antitumor activity of acid labile transferring and albumindoxorubicin conjugates in in vitro and in vivo human tumor xenograft models. European Journal of Cancer, 33, S175.

    Article  Google Scholar 

  4. Gutmann, O., Kuehlewein, R., Reinbold, S., & Niekrawietz, R. (2005). Fast and reliable protein microarray production by a new drop-in-drop technique. Lab on a Chip, 5, 675–681.

    Article  CAS  Google Scholar 

  5. Mircean, C., Shmulevich, I., Cogdell, D., Choi, W., Jia, Y., Tabus, I., Hamilton, S. R., & Zhang, W. (2005). Robust estimation of protein expression ratios with lysate microarray technology. Bioinformatics, 21, 1935–1942.

    Article  CAS  Google Scholar 

  6. Willison, H. J., Townson, K., Veitch, J., Boffey, J., Isaacs, N., Andersen, S. M., Ling, P. Z. C.-C., & Bundle, D. R. (2004). Synthetic disialylgalactose immunoadsorbents deplete anti-GQ1b antibodies from autoimmune neuropathy sera. Brain, 127, 680–691.

    Article  Google Scholar 

  7. Mirapurkar, S., Nagvekar, U. H., & Sivaprasad, N. (2007). Polyreactivity of monoclonal antibodies produced against thyroid stimulating hormone (hTSH). Journal of Immunoassay Immunochemistry, 28, 119–126.

    Article  CAS  Google Scholar 

  8. Studentsov, Y., Schiffman, M., Strickler, H. D., Ho, G. Y. F., Pang, Y. Y. S., Schiller, J., Herrero, R., & Burk, D. (2002). Enhanced enzyme-linked immunosorbent assay for detection of antibodies to virus-like particles of human papillomavirus. Journal of Clinical Microbiology, 40, 1755–1760.

    Article  CAS  Google Scholar 

  9. Qiu, B., Stefanos, S., Ma, J., Lalloo, A., Perry, B. A., Leibowitz, M. J., Sinko, P. J., & Stein, S. (2003). A hydrogel prepared by in situ cross-linking of a thiolcontaining poly (ethylene glycol)-based copolymer: a new biomaterial for protein delivery. Biomaterials, 24, 11–18.

    Article  Google Scholar 

  10. Chia, H. H., Yang, Y. Y., Chung, T. S., Ng, S., & Heller, J. (2001). Auto-catalyzed poly (ortho ester) microspheres: a study of their erosion and release mechanism. Journal of Controlled Release, 75, 11–25.

    Article  CAS  Google Scholar 

  11. Kyselova, Z., Rackova, L., & Stefek, M. (2003). Pyridoindole antioxidant stobadine protected bovine serum albumin against the hydroxyl radical mediated crosslinking in vitro. Archives of Gerontology and Geriatrics, 36, 221–229.

    Article  CAS  Google Scholar 

  12. Hoshi, S., Saito, N., Kusanagi, K., Ihara, T., & Ueda, S. (1998). Adjuvant effects of fluoride on oral immunization of chickens. Veterinary Immunology and Immunopathology, 63, 253–263.

    Article  CAS  Google Scholar 

  13. Sprott, G. D., Tolson, D. L., & Patel, G. B. (1997). Archaeosomes as novel antigen delivery systems. FEMS Microbiology Letters, 15, 17–22.

    Article  Google Scholar 

  14. Ramaldes, G. A., Deverre, J. R., Grognet, J. M., Puisieux, F., & Fattal, E. (1996). Use of an enzyme immunoassay for the evaluation of entrapment efficiency and in vitro stability in intestinal fluids of liposomal bovine serum albumin. International Journal of Pharmaceutics, 143, 1–11.

    Article  CAS  Google Scholar 

  15. Hilleman, M. R. (2000). Vaccines in historic evolution and perspective: a narrative of vaccine discoveries. Vaccine, 18, 1436–1447.

    Article  CAS  Google Scholar 

  16. Ada, G. (2003). Overview of vaccines. Methods in Molecular Medicine, 87, 1–17.

    CAS  Google Scholar 

  17. Pastorret, P. P., Brochier, B., & Denis, M. (1995). Environmental safety assessment of vaccines derived from biotechnology. Revue Scientifique et Technique, 14, 963–977.

    Google Scholar 

  18. Huang, C. Y., Liang, C. M., Li, C. Chu, Mei Liang, S. (2009). Albumin fibrillization induces apoptosis via integrin/FAK/Akt pathway. BMC Biotechnology, 9, 1–12.

    Google Scholar 

  19. Khamehchian, S., Madani, R., Golchinfar, F., & Taghavian, M. (2008). Development of a sandwich enzyme-linked immunosorbent assay (ELISA) for determining of bovine serum albumin (BSA) in trivalent measles–mumps–rubella (MMR) vaccines. Human Vaccines, 4, 375–378.

    Article  CAS  Google Scholar 

  20. Fiocchi, A., Restani, P., Riva, E., Qualizza, R., Bruni, P., & Restelli, A. R. (1995). Meat allergy: 1—specific IgE to BSA and OSA in atopic, beef sensitive children. Journal of the American College of Nutrition, 14, 239–244.

    CAS  Google Scholar 

  21. Fiocchi, A., Restani, P., Riva, E., Mirri, G. P., Santini, I., & Bernardo, L. (1998). Heat treatment modifies the allergenicity of beef and bovine serum albumin. Allergy, 53, 798–802.

    Article  CAS  Google Scholar 

  22. Martelli, A., Chiara, A., Corvo, M., Restani, P., & Fiocchi, A. (2002). Beef allergy in children with cow’s milk allergy; cow’s milk allergy in children with beef allergy. Annals of Allergy, Asthma & Immunology, 89, 38–43.

    Article  Google Scholar 

  23. Vicente, S. J., Caballero, M. L., Rodriguez, P. R., Carretero, P., Perez, R., & Blanco, J. G. (2007). Sensitization to serum albumins in children allergic to cow’s milk and epithelia. Pediatric Allergy and Immunology, 18, 503–507.

    Article  Google Scholar 

  24. Tanabc, S., Kobayashi, Y., Takahata, Y., Morimatsu, F., Shibata, R., & Nishimura, T. (2002). Some human B and T cell epitopes of bovine serum albumin, the major beef allergen. Biochemical and Biophysical Research Communications, 293, 1348–1353.

    Article  Google Scholar 

  25. Day, M. J. (2007). Vaccine safety in the neonatal period. Journal of Comparative Pathology, 137, 51–56.

    Article  Google Scholar 

  26. Ter Avest, A. R., Van Steenis, G., & Osterhaus, A. D. (1987). A comparison of an enzyme-linked immunosorbent assay and counter current electrophoresis for the detection of bovine serum albumin in virus vaccines. Journal of Biological Standardization, 15, 245–250.

    Article  Google Scholar 

  27. Yu, Y., Lai, Y., Zheng, X. L., Wu, J. Z., Long, Z. Y., & Liang, C. S. (2007). Synthesis of functionalized CdTe/CdS QDs for spectrofluorimetric detection of BSA. Spectrochim Acta Part A: Molecular and Biomolecular Spectroscopy, 68, 1356–1361.

    Article  Google Scholar 

  28. Rubtosova, M. J., Wittman, C., Egorov, A. M., & Schmid, R. D. (1997). Thermoalkalophilic lipase of Bacillus thermocatenulatus: large-scale production, purification and properties: aggregation behaviour and its effect on activity. Journal of Biotechnology, 56, 89–102.

    Google Scholar 

  29. Dzgoev, A. B., Gazaryan, I. G., Lagrimini, L. M., Ramanathan, K., & Danielsson, B. (1999). High-sensitivity assay for pesticide using a peroxidase as chemiluminescent label. Analytica Chimica Acta, 71, 5258–5261.

    CAS  Google Scholar 

  30. Tan, X. J., Song, Z. H., Chen, D. H., & Wang, Z. M. (2011). Study on the chemiluminescence behavior of bovine serum albumin with luminol and its analytical application. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 79, 232–235.

    Article  CAS  Google Scholar 

  31. Zhang, K., Song, C. J., Li, Q., Li, Y. M., Sun, Y. J., Yang, K., & Jin, B. Q. (2010). The establishment of a highly sensitive ELISA for detecting bovine serum albumin (BSA) based on a specific pair of monoclonal antibodies (nAb) and its application in vaccine quality control. Human Vaccines, 6, 1–7.

    Article  Google Scholar 

  32. Liang, S. X., Lia, H. F., & Lin, J. M. (2008). Reaction mechanism of surfactant-sensitized chemiluminescence of bis(2,4,6- trichlorophyenyl) oxalate and hydrogen peroxide induced by gold nanoparticles. Luminescence, 23, 381–385.

    Article  CAS  Google Scholar 

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Acknowledgement

This work was supported financially by the Fundamental Research Funds for the Central Universities (Program No. GK20091004), the National High–tech R&D Program (863 Program) of China (No. 2006AA02A237), and the National Natural Science Foundation of China (No. 30872371).

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Authors and Affiliations

  1. Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Materials Science, Shaanxi Normal University, Xi’an, 710062, China

    Pan Xue, Zhujun Zhang, Yun Li, Feng Liu, Xiaoming Zhang & Aihua Fu

  2. Department of Immunology, Fourth Military Medical University, Xi’an, 710032, China

    Kui Zhang, Yuanjie Sun, Chaojun Song, Boquan Jin & Kun Yang

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  1. Pan Xue
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  2. Kui Zhang
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  3. Zhujun Zhang
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  4. Yun Li
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  5. Feng Liu
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  6. Yuanjie Sun
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  7. Xiaoming Zhang
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  8. Chaojun Song
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  9. Aihua Fu
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Correspondence to Zhujun Zhang or Kun Yang.

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Pan Xue and Kui Zhang contributed equally to this work.

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Xue, P., Zhang, K., Zhang, Z. et al. Highly Sensitive Chemiluminescent Analysis of Residual Bovine Serum Albumin (BSA) Based on a Pair of Specific Monoclonal Antibodies and Peroxyoxalate–glyoxaline–PHPPA Dimer Chemiluminescent System in Vaccines. Appl Biochem Biotechnol 166, 1604–1614 (2012). https://doi.org/10.1007/s12010-012-9567-1

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