Desarrollo de superficies modificadas oro - anticuerpo anti-PSPB para su uso como biosensor en la detección de preñez bovina

Authors

  • Justo Laíz Unidad de Bioquímica Analítica, Centro de Investigaciones Nucleares (CIN), Facultad de Ciencias , Universidad de la República, Uruguay
  • Karen Keel Unidad de Bioquímica Analítica, Centro de Investigaciones Nucleares (CIN), Facultad de Ciencias , Universidad de la República, Uruguay, Departamento de Aguas y Productos Químicos, Laboratorio Tecnológico del Uruguay (LATU), Uruguay
  • Juan Pablo Tosar Unidad de Bioquímica Analítica, Centro de Investigaciones Nucleares (CIN), Facultad de Ciencias , Universidad de la República, Uruguay

DOI:

https://doi.org/10.26461/05.04

Abstract

Due to big importance of early clinical diagnosis of pregnancy in cattle, the aim of this work was the development of an ampero- metric biosensor which enables protein quantification of pregnancy specific protein B in biological fluids. To achieve this goal Mercato Undecanoic Acid was immobilized on the gold surface, with anti-PSPB and Horseradish Peroxidase. The immobiliza- tion was verified by electrochemical and non electrochemical methods, such as cyclic voltammetry and contact angle measure-ments respectively.The electrochemical performance of the electrode was studied by cyclic voltammetry and constant potential amperometry, to establish the relationship between the change in cathode current (the quinone reduction potential) and the concentration of PSPB. Due to the formation of large antigen antibody complexes which limit the access of the enzyme’s specific substrate to its active site, the cathodic current would decrease as was observed.The developed gold - anti-PSPB - HRP electrode showed a lineal dependence on the concentration of PSPB at potential of -200 mV (vs Ag/AgCl), both by cyclic voltammetry and constant potential amperometry. Therefore this study could be use for the design of an amperometric biosensor which allows the detection of bovine pregnancy at the earlier stages.

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References

BUTLER, J.; HAMILTON, W.; SASSER, R.; RUDER, C.; HASS, G.; WILLIAMS, R. Detection and partial characterization of two bovine pregnancyspecific proteins. En: Biology of Reproduction. 1982, 26(5):925-933.

CHAKI, N.; VIJAYAMOHANAN, K. Self-assembled monolayers as a tunable platform for biosensor applications. En: Biosensors and Bioelectronics. 2002, 17(1-2):1-12.

DANCZYK, R.; KRIEDER, B.; NORTH, A.; WEBSTER, T.; HOGENESCH, H.; RUNDELL, A. Comparison of antibody functionality using different immobilization methods. En: Biotechnology and Bioengineering. 2003, 84(2):215-223.

DEL VECCHIO, R.; SUTHERLAND, W.; SASSER, R. Effect of pregnancy-specific protein B on luteal cell progesterone, prostaglandin, and oxytocin production during two stages of the bovines estrous cycle. En: Journal of Animals Science. 1995, 73:2662-2668.

EVTUGYN, G.; MINGALEVA, A.; BUDNIKOV, H.; STOIKOVA, E.; VINTER, V.; EREMIN, S. Affinity biosensors based on disposable screen-printed electrodes modified with DNA. En: Analytica Chimica Acta. 2003, 479(2):125-134.

FU, Y.; YUAN, R.; TANG, D.; CHAI, Y. XU, L. Study on the immobilization of anti-IgG on Au-colloid modified gold electrode via. En: Colloids and Surfaces B: Biointerfaces. 2005, 40(1):61-66.

GHINDILI, A.; ATANASOV, P.; WIKINS, M. y WIKINS, E. Immunosensors: electrochemical sensing and other engineering approaches. En: Biosensors and Bioelectronics. 1998, 13:113-131.

GILEADI, E. Electrode kinetics for chemists, chemical engineers, and material scientists. New York: VCH Publishers, 1993. ISBN: 0-471-18858-1.

KIRACOFE, G., WRIGHT, J.; SCHALLES, R.; RUDER, C.; PARISH, S. Y SASSER R. Pregnancy-specific protein B in serum of postpartum beef cows. En: Journal of Animal Science. 1993, (71):2199-2205.

LAGES, C. Implementación de la técnica de ángulo de contacto para el estudio de interfases. Montevideo: Facultad de Ciencias, 2006. (Tesis de grado)

MELLO, L.; KUBOTA, L. Review of the use of biosensors as analytical tools in the food and drink industries. En: Food Chemistry. 2002, 77(2):237-256.

MORGAN, C.; NEWMAN, D.; PRICE, C. Immunosensors: technology and opportunities in laboratory medicine. En: Clinical Chemistry. 1996, (42):193-209.

ROPSTAD, E.; JOHANSEN, O.; KING, C.; DAHL, E.; ALBON, S.; LANGVATN, R.; IRVINE, R.; HALVORSEN, O.; SASSER, R. Comparison of plasma progesterone, transrectal ultrasound and pregnancy specific proteins (PSPB) used for pregnancy diagnosis in reindeer. En: Acta Vet. Scand. 1999, 40(2):151-162.

RUDER C.; STELLFLUG, J.; DAHMEN J.; SASSER, R. Detection of pregnancy in sheep of sera for pregnancy-specific protein B. En: Theriogenology. 1988, 29(4):905-911.

SASSER, R.; RUDER, C.; IVANI, K.; BUTLER, J. y HAMILTON W. Detection of pregnancy by radioimmunoassay of a novel pregnancy-specific protein in serum of cows and a profile of serum concentrations during gestation. En: Biology of Reproduction. 1986, 35:936-942.

SCHREIBER, F. Structure and growth of self-assembling monolayers. En: Progress in Surfeces Science. 2000, 65(5):151-256.

SEMAMBO, D.; AYLIFFET, R.; BOYD, J.; Y TAYLOR, D. Ultrasonographic study of early embryonic loss induced by Actinomyces pyogenes in cattle. En: Veterinarian Record. 1992, 131:7-12.

SHAOJUN, D; JINGHONG, L. Self-assembled monolayers of thiols on gold electrodes for bioelectrochemistry and biosensors. En: Bioelectrochemistry and Bioenergetics. 1997, 42(1):7-13.

SUN, T.; FENG, L.; GAO, X.; JIANG, L. Bioinspired surfaces with special wettability. En: Acc. Chemycal. 2005, 38(8):644-352

WANG, M.; WANG, L.; WANG, G.; JI, X.; BIE Y.; LI, T.; GONG, S. Y LI, J. Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reactions. En: Biosensors and Bioelectronics. 2004, 19(6):575-582.

WANG Z.; VIANA, A.; JIN, G.; ABRANTES, L. Immunosensor interface based on physical and chemical immunoglobulin G adsorption onto mixed self-assembled monolayers. En; Bioelectrochemestry. 2006, 69(2):180-186.

How to Cite

Laíz, J., Keel, K., & Tosar, J. P. (2011). Desarrollo de superficies modificadas oro - anticuerpo anti-PSPB para su uso como biosensor en la detección de preñez bovina. INNOTEC, (5 ene-dic), 29–33. https://doi.org/10.26461/05.04

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