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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:ali="http://www.niso.org/schemas/ali/1.0/" article-type="research-article" dtd-version="1.2" xml:lang="en"><front><journal-meta><journal-id journal-id-type="publisher-id">Acta Naturae</journal-id><journal-title-group><journal-title xml:lang="en">Acta Naturae</journal-title><trans-title-group xml:lang="ru"><trans-title>Acta Naturae</trans-title></trans-title-group></journal-title-group><issn publication-format="print">2075-8251</issn><publisher><publisher-name xml:lang="en">Acta Naturae Ltd</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="publisher-id">10697</article-id><article-id pub-id-type="doi">10.32607/20758251-2011-3-3-85-92</article-id><article-categories><subj-group subj-group-type="toc-heading" xml:lang="en"><subject>Articles</subject></subj-group><subj-group subj-group-type="toc-heading" xml:lang="ru"><subject>Статьи</subject></subj-group><subj-group subj-group-type="article-type"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title xml:lang="en">Recombinant Production of Horseradish Peroxidase Conjugates with Fab Antibodies in Pichia pastoris for Analytical Applications</article-title><trans-title-group xml:lang="ru"><trans-title>Recombinant Production of Horseradish Peroxidase Conjugates with Fab Antibodies in Pichia pastoris for Analytical Applications</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author"><name><surname>Koliasnikov</surname><given-names>O V</given-names></name><bio xml:lang="en"><p>Department of Chemistry</p></bio><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><name><surname>Grigorenko</surname><given-names>V G</given-names></name><bio xml:lang="en"><p>Department of Chemistry</p></bio><email>vitaly@immunotek.ru</email><xref ref-type="aff" rid="aff2"/></contrib><contrib contrib-type="author"><name><surname>Egorov</surname><given-names>A M</given-names></name><bio xml:lang="en"><p>Department of Chemistry</p></bio><xref ref-type="aff" rid="aff2"/></contrib><contrib contrib-type="author"><name><surname>Lange</surname><given-names>S</given-names></name><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><name><surname>Schmid</surname><given-names>R D</given-names></name><xref ref-type="aff" rid="aff3"/></contrib></contrib-group><aff-alternatives id="aff1"><aff><institution xml:lang="en">Kolmogorov Advanced Education and Science Center, Lomonosov Moscow State University</institution></aff><aff><institution xml:lang="ru"></institution></aff></aff-alternatives><aff-alternatives id="aff2"><aff><institution xml:lang="en">Lomonosov Moscow State University</institution></aff><aff><institution xml:lang="ru"></institution></aff></aff-alternatives><aff-alternatives id="aff3"><aff><institution xml:lang="en">Institute of Technical Biochemistry, University of Stuttgart</institution></aff><aff><institution xml:lang="ru"></institution></aff></aff-alternatives><pub-date date-type="pub" iso-8601-date="2011-09-15" publication-format="electronic"><day>15</day><month>09</month><year>2011</year></pub-date><volume>3</volume><issue>3</issue><issue-title xml:lang="en">VOL 3, NO3 (2011)</issue-title><issue-title xml:lang="ru">ТОМ 3, №3 (2011)</issue-title><fpage>85</fpage><lpage>92</lpage><history><date date-type="received" iso-8601-date="2020-01-17"><day>17</day><month>01</month><year>2020</year></date></history><permissions><copyright-statement xml:lang="en">Copyright ©; 2011, Koliasnikov O.V., Grigorenko V.G., Egorov A.M., Lange S., Schmid R.D.</copyright-statement><copyright-statement xml:lang="ru">Copyright ©; 2011, Koliasnikov O.V., Grigorenko V.G., Egorov A.M., Lange S., Schmid R.D.</copyright-statement><copyright-year>2011</copyright-year><copyright-holder xml:lang="en">Koliasnikov O.V., Grigorenko V.G., Egorov A.M., Lange S., Schmid R.D.</copyright-holder><copyright-holder xml:lang="ru">Koliasnikov O.V., Grigorenko V.G., Egorov A.M., Lange S., Schmid R.D.</copyright-holder><ali:free_to_read xmlns:ali="http://www.niso.org/schemas/ali/1.0/"/><license><ali:license_ref xmlns:ali="http://www.niso.org/schemas/ali/1.0/">https://creativecommons.org/licenses/by/4.0</ali:license_ref></license></permissions><self-uri xlink:href="https://actanaturae.ru/2075-8251/article/view/10697">https://actanaturae.ru/2075-8251/article/view/10697</self-uri><abstract xml:lang="en"><p/></abstract><trans-abstract xml:lang="ru"><p>Recombinant immunoconjugates of marker enzymes with antigens or antibodies present considerably more advantages than those obtained by conventional methods of chemical synthesis; i.e. they are homogeneous, have a strictly determined stoichiometry, and retain the functional activity of both a marker protein and an antigen/antibody. Based on the pPICZαB shuttle vector, we first managed to obtain a recombinant conjugate of key marker enzyme horseradish peroxidase (HRP) with Fab fragments of antibodies against atrazine. The resulting genetic construction allows us to switch to any other antibody sequence, via the simple re-cloning of variable parts and an additional reporter enzyme. Conjugates were successfully produced in the Pichia pastoris methylotrophic yeast expression system. The target activity of the conjugates (both enzymatic and antigen-binding) has been demonstrated by ELISA method.</p></trans-abstract><kwd-group xml:lang="en"><kwd>horseradish peroxidase</kwd><kwd>antibodies</kwd><kwd>recombinant conjugates</kwd><kwd>Pichia pastoris expression. ABBREVIATIONS HRP - horseradish peroxidase</kwd><kwd>ELISA - enzyme-linked immunosorbent assay</kwd><kwd>BSA - bovine serum albumin</kwd><kwd>PCR - polymerase chain reaction</kwd><kwd>TMB - 3,3’,5,5’-tetramethylbenzidine</kwd><kwd>ABTS - 2,2’-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)</kwd></kwd-group></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Rau D., Kramer K., Hock B. // J. Immunoassay Immunochem. 2002. V. 23. № 2. Р. 129-143.</mixed-citation></ref><ref id="B2"><label>2.</label><mixed-citation>Tachibana H., Takekoshi M., Cheng X.J., Nakata Y., Takeuchi T., Ihara S. // Clin. Diagn. Lab. Immunol. 2004. V. 11. №1. Р. 216-218.</mixed-citation></ref><ref id="B3"><label>3.</label><mixed-citation>Mousli M., Turki I., Kharmachi H., Saadi M., Dellagi K. // J. Virol. Meth. 2007. V. 146. №1-2. 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