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Preparation of monospecific antibodies against isoform 2 of translation elongation factor 1A (eEF1A2)

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Abstract

Amino acid sequences of eukaryotic translation elongation factor isoform 1 (eEF1A1) and 2 (eEF1A2) were compared and two peptide fragments of eEF1A2 were chosen as linear antigenic determinants for generation of monospecific antipeptide antibodies. Synthesized peptides corresponded to the selected peptide fragments were conjugated to bovine serum albumin (BSA) and used for immunizations of mice. Antibodies, produced against the eEF1A2 fragment 330–343 conjugated to BSA, specifically recognized this isoform in the native and partially denatured states but did not interact with the eEF1A1 isoform. It was shown that these monospecific anti-eEF1A2 antibodies could be employed for eEF1A2 detection both by enzyme-linked immunosorbent assay and by immunoblotting.

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Authors and Affiliations

  1. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, ul. Pogodinskaya 10, Moscow, 119121, Russia

    E. F. Kolesanova, T. E. Farafonova, E. Yu. Aleshina & N. V. Pyndyk

  2. State Key Laboratory of Molecular and Cell Biology, Institute of Molecular Biology and Genetics of the National Academy of Sciences, ul. akad. Zabolotnogo 150, Kiev, 03143, Ukraine

    M. V. Veremieva, A. V. Novosylnaya, M. I. Kovalenko, V. F. Shalak & B. S. Negrutskii

Authors
  1. E. F. Kolesanova
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  2. T. E. Farafonova
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  3. E. Yu. Aleshina
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  4. N. V. Pyndyk
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  5. M. V. Veremieva
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  6. A. V. Novosylnaya
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  7. M. I. Kovalenko
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  8. V. F. Shalak
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  9. B. S. Negrutskii
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Correspondence to E. F. Kolesanova.

Additional information

Original Russian Text © E.F. Kolesanova, T.E. Farafonova, E.Yu. Aleshina, N.V. Pyndyk, M.V. Veremieva, A.V. Novosylnaya, M.I. Kovalenko, V.F. Shalak, B.S. Negrutskii, 2013, published in Biomeditsinskaya Khimiya.

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Kolesanova, E.F., Farafonova, T.E., Aleshina, E.Y. et al. Preparation of monospecific antibodies against isoform 2 of translation elongation factor 1A (eEF1A2). Biochem. Moscow Suppl. Ser. B 7, 62–69 (2013). https://doi.org/10.1134/S1990750813010083

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  • DOI: https://doi.org/10.1134/S1990750813010083

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