Preparation of human tau exon-2- and -10-specific monoclonal antibodies for the recognition of brain tau proteins in various mammals

  • Authors:
    • Cao Chen
    • Yan Lv
    • Qi Shi
    • Bao-Yun Zhang
    • Li-Na Chen
    • Kang Xiao
    • Jing Sun
    • Xiao-Ping Dong
  • View Affiliations

  • Published online on: June 3, 2015     https://doi.org/10.3892/ijmm.2015.2235
  • Pages: 455-462
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Abstract

The aggregations of tau protein in brain tissue have been described in a large number of neurodegenerative diseases; however, due to the lack of tau isoform- or exon-specific antibodies, the exact situations under which various brain tau isoforms can be found and their exact contributions during disease progression remain unknown. Therefore, in this study, we prepared tau exon-specific monoclonal antibodies (mAbs) that recognize different mammalian tau isoforms. Briefly, 3 Balb/c mice were separately immunized (3 mice per antigen) with the recombinant GST-fusion proteins, GST-tE2 and GST-tE10. Two hybridoma cell lines, 4A8 and 3E12, secreting antibodies against human tau exon-2 and -10 were established using the hybridoma technique. The sensitivity and specificity of the prepared mAbs were evaluated using indirect ELISA and western blot analysis. The ability of the prepared mAbs, 4A8 and 3E12, to recognize endogenous tau protein in the brain tissues of various mammals was estimated by immunoprecipitation. Based on the results of various verification methods, we found that the prepared mAbs, 4A8 and 3E12, not only specifically reacted with the individual recombinant GST tau exon fusion proteins, but also correctly recognized the recombinant human tau isoforms containing respective exon sequences, as shown by western blot analysis. Furthermore, western blot analysis and immunoprecipitation assays verified that the mAbs, 4A8 and 3E12, recognized endogenous tau proteins in human brain tissue, as well as tau proteins in a series of mammalian tissues, including goat, bovine, rabbit, hamster and mouse. Thus, in the present study, using the hybridoma technique, we successfully prepared the mAbs, 4A8 against tau exon-2 and 3E12 against tau exon-10, which provide useful tools for determining potential alternations of tau isoforms in neurodegenerative diseases.
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August-2015
Volume 36 Issue 2

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Chen C, Lv Y, Shi Q, Zhang B, Chen L, Xiao K, Sun J and Dong X: Preparation of human tau exon-2- and -10-specific monoclonal antibodies for the recognition of brain tau proteins in various mammals. Int J Mol Med 36: 455-462, 2015.
APA
Chen, C., Lv, Y., Shi, Q., Zhang, B., Chen, L., Xiao, K. ... Dong, X. (2015). Preparation of human tau exon-2- and -10-specific monoclonal antibodies for the recognition of brain tau proteins in various mammals. International Journal of Molecular Medicine, 36, 455-462. https://doi.org/10.3892/ijmm.2015.2235
MLA
Chen, C., Lv, Y., Shi, Q., Zhang, B., Chen, L., Xiao, K., Sun, J., Dong, X."Preparation of human tau exon-2- and -10-specific monoclonal antibodies for the recognition of brain tau proteins in various mammals". International Journal of Molecular Medicine 36.2 (2015): 455-462.
Chicago
Chen, C., Lv, Y., Shi, Q., Zhang, B., Chen, L., Xiao, K., Sun, J., Dong, X."Preparation of human tau exon-2- and -10-specific monoclonal antibodies for the recognition of brain tau proteins in various mammals". International Journal of Molecular Medicine 36, no. 2 (2015): 455-462. https://doi.org/10.3892/ijmm.2015.2235