Open AccessA quantitative method for determination of PPDK concentration in miscanthus leaves
Author(s) -
Meshcheryakova Irina A.,
Bannikova Svetlana V.,
Rozanov Aleksey S.,
Demidova Elisaveta V.,
Demidov Evgeniy A.,
Goryachkovskaya Tatia.,
Burmakitalya V.,
Shekhovtsov Sergey V.,
Bryanskaya Alla V.,
Kolchanov Nikolay A.,
Peltek Sergey E.
Publication year - 2017
Publication title -
gcb bioenergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.378
H-Index - 63
eISSN - 1757-1707
pISSN - 1757-1693
DOI - 10.1111/gcbb.12361
Subject(s) - miscanthus , monoclonal antibody , chemistry , mass spectrometry , western blot , chromatography , affinity chromatography , microbiology and biotechnology , antibody , biochemistry , biology , gene , enzyme , bioenergy , genetics , biofuel
In this study, we used ELISA for quantification of PPDK in photosynthesizing leaves of miscanthus. We cloned a fragment of the gene encoding PPDK , purified the resulting protein by affinity chromatography, identified it using MALDI mass spectrometry, and obtained monoclonal antibodies by immunizing BALB /c mice. Selectivity of monoclonal antibodies was assessed by Western blot using the protein extracts of Soranovskii. The presence of PPDK was again verified by MALDI mass spectrometry. Therefore, we developed and tested the method for determining PPDK quantity in miscanthus using ELISA .