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

Kolesanova E.F.1 , Farafonova T.E.1, Aleshina E.Yu.1, Pyndyk N.V.1, Veremieva M.V.2, Novosylna O.V.2, Kovalenko M.I.2, Shalak V.F.2, Negrutskii B.S.2

1. Orekhovich Institute of Biomedical Chemistry of the Russian Academy of Medical Sciences
2. State Key Laboratory of Molecular and Cell Biology, Institute of Molecular Biology and Genetics of the National Academy of Sciences
Section: Experimental Study
DOI: 10.18097/PBMC20146001051      PubMed Id: 24749247
Year: 2014  Volume: 60  Issue: 1  Pages: 51-62
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. Selected peptides were synthesized, conjugated to bovine serum albumin (BSA) and used for mice immunizations. 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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Keywords: translation elongation factor 1A, antipeptide antibodies, peptide synthesis, enzyme-linked immunosorbent assay, immunoblotting

Kolesanova, E. F., Farafonova, T. E., Aleshina, E. Yu., Pyndyk, N. V., Veremieva, M. V., Novosylna, O. V., Kovalenko, M. I., Shalak, V. F., Negrutskii, B. S. (2014). Preparation of monospecific antibodies against isoform 2 of translation elongation factor 1A (eEF1A2). Biomeditsinskaya Khimiya, 60(1), 51-62.
This paper is also available as the English translation: 10.1134/S1990750813010083
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