Share This Article:

Preparation of monospecific anti-PAG antibodies for cattle pregnancy detection: use of synthetic peptides to improve specificity

Abstract Full-Text HTML Download Download as PDF (Size:774KB) PP. 85-88
DOI: 10.4236/abb.2011.22013    5,090 Downloads   10,076 Views   Citations

ABSTRACT

Immunological methods involving pregnancy associ-ated glycoproteins (PAG) are used for cattle preg-nancy detection. The faults of these methods could be overcomed by using antibodies specific for each member of the PAG family. In order to differentiate between very similar proteins, preparation of anti-bodies specific for peptides is a method of choice. In this work, we summarize a series of considerations regarding peptide design and choose free access NCBI, Antigenicity Plot, EMBOSS Antigenic and Expasy tools to apply them. We design peptides spe-cific for different reported PAG members and obtain the corresponding polyclonal antibodies for five of them.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Álvarez, J. , Teijeiro, J. and Marini, P. (2011) Preparation of monospecific anti-PAG antibodies for cattle pregnancy detection: use of synthetic peptides to improve specificity. Advances in Bioscience and Biotechnology, 2, 85-88. doi: 10.4236/abb.2011.22013.

References

[1] Piechotta, M., Bollwein, J., Friedrich, M., Heilkenbrinker, T., Passavant, C., Branen, J., Sasser, G., Hoedemaker, M. and Bollwein, H. (2011) Comparison of commercial ELISA blood tests for early pregnancy detection in dairy cows. Journal of Reproduction and Development, 57, 72-75.
[2] Green, J.A., Xie, S., Quan, X., Bao, B., Gan, X., Mathialagan, N., Beckers, J.F. and Roberts, R.M. (2000) Preg- nancy-associated bovine and ovine glycoproteins exhibit spatially and temporally distinct expression patterns during pregnancy. Biology of Reproduction, 62, 1624-1631. doi:10.1095/biolreprod62.6.1624
[3] Ayad, A., Sousa, N.M., Sulon, J., Hornick, J.L., Iguer- Ouada, M. and Beckers, J.F. (2009) Correlation of five radioimmunoassay systems for measurement of bovine plasma pregnancy-associated glycoprotein concentrations at early pregnancy period. Research in Veterinary Science, 86, 377-382. doi:10.1016/j.rvsc.2008.10.003
[4] Whitlock, B.K. and Maxwell, H.S. (2008) Pregnancy- associated glycoproteins and pregnancy wastage in cattle. Theriogenology, 70, 550-559. doi:10.1016/j.theriogenology.2008.05.003
[5] Green, J.C., Volkmann, D.H., Poock, S.E., McGrath, M.F., Ehrhardt, M., Moseley, A.E. and Lucy, M.C. (2009) Technical note: A rapid enzyme-linked immunosorbent assay blood test for pregnancy in dairy and beef cattle. Journal of Dairy Science, 92, 3819-3824. doi:10.3168/jds.2009-2120.
[6] Saccodossi, N., De Simone, E. and Leoni, J. (2006) A novel and easy method for the production of recombinant peptides for use in the generation of monospecific antisera against Lama glama IgG2b and IgG2c subclasses. Journal of Immunological Methods, 313, 214-218. doi:10.1016/j.jim.2006.05.005
[7] Harlow, E. and Lane, D. (1988) Antibodies, a laboratory manual. Cold Spring Harbor Laboratory.
[8] NCBI database. http://www.ncbi.nlm.nih.gov/
[9] Corpet, MultAlin5.4.1. (1988) Internet available at http://bioinfo.genotoul.fr/multalin/multalin.htlm
[10] Hopp and Woods (1981) Antigenicity Plot. Internet http://www.bioinformatics.org/JaMBW/3/1/7/
[11] Kolaskar and Tongaonkar (1990) EMBOSS Antigenic. http://liv.bmc.uu.se/cgi-bin/emboss/antigenic.
[12] Hughes, A.L., Green, J.A., Garbayo, J.M. and Roberts, R.M. (2000) Adaptive diversification within a large family of recently duplicated, placentally expressed genes. Proceedings of the National Academy of Sciences USA, 97, 3319-3323. doi:10.1073/pnas.050002797
[13] Expasy tools. http://ca.expasy.org/tools/
[14] Peptide Calculator, Sigma-Aldrich. Internet available at: http://www.sigma-genosys.com/calc/pepCalc.asp
[15] Xie, S., Green, J. and Roberts, R.M. (1988) Expression of multiple genes for pregnancy-associated glycoproteins in the sheep placenta. Advances in Experimental Medicine and Biology, 436, 195-200.

  
comments powered by Disqus

Copyright © 2019 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.