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Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol
This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for Escherichia coli Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parame...
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| Published in: | Russian journal of bioorganic chemistry 2016, Vol.42 (1), p.22-28 |
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| Main Authors: | , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c321t-54849766fc24abeddd8c0aa3944d1d40bd7fffe260532235dbbb71c5c75fc7183 |
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| cites | cdi_FETCH-LOGICAL-c321t-54849766fc24abeddd8c0aa3944d1d40bd7fffe260532235dbbb71c5c75fc7183 |
| container_end_page | 28 |
| container_issue | 1 |
| container_start_page | 22 |
| container_title | Russian journal of bioorganic chemistry |
| container_volume | 42 |
| creator | Dormeshkin, D. O. Kuprienko, O. S. Svirid, A. V. Gilep, A. A. Sviridov, O. V. Usanov, S. A. |
| description | This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for
Escherichia coli
Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parameters of interaction of these antibody fragments with cortisol-BSA complex were measured via the biolayer interferometry method. An equilibrium dissociation constant of this interaction K
D
is 5.48 × 10
–10
M. The interaction is reversible and it could be competitively inhibited by free cortisol addition. These results could be used in generating of immunoassays for quantitative cortisol determinations. The in vivo biotinylation system under review is universal and suitable for expression of any biotinylated Fab fragments in the
E. coli
system. |
| doi_str_mv | 10.1134/S1068162016010039 |
| format | article |
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Escherichia coli
Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parameters of interaction of these antibody fragments with cortisol-BSA complex were measured via the biolayer interferometry method. An equilibrium dissociation constant of this interaction K
D
is 5.48 × 10
–10
M. The interaction is reversible and it could be competitively inhibited by free cortisol addition. These results could be used in generating of immunoassays for quantitative cortisol determinations. The in vivo biotinylation system under review is universal and suitable for expression of any biotinylated Fab fragments in the
E. coli
system.</description><identifier>ISSN: 1068-1620</identifier><identifier>EISSN: 1608-330X</identifier><identifier>DOI: 10.1134/S1068162016010039</identifier><language>eng</language><publisher>Moscow: Pleiades Publishing</publisher><subject>Biochemistry ; Biomedical and Life Sciences ; Biomedicine ; Bioorganic Chemistry ; Escherichia coli ; Life Sciences ; Organic Chemistry</subject><ispartof>Russian journal of bioorganic chemistry, 2016, Vol.42 (1), p.22-28</ispartof><rights>Pleiades Publishing, Ltd. 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c321t-54849766fc24abeddd8c0aa3944d1d40bd7fffe260532235dbbb71c5c75fc7183</citedby><cites>FETCH-LOGICAL-c321t-54849766fc24abeddd8c0aa3944d1d40bd7fffe260532235dbbb71c5c75fc7183</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids></links><search><creatorcontrib>Dormeshkin, D. O.</creatorcontrib><creatorcontrib>Kuprienko, O. S.</creatorcontrib><creatorcontrib>Svirid, A. V.</creatorcontrib><creatorcontrib>Gilep, A. A.</creatorcontrib><creatorcontrib>Sviridov, O. V.</creatorcontrib><creatorcontrib>Usanov, S. A.</creatorcontrib><title>Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol</title><title>Russian journal of bioorganic chemistry</title><addtitle>Russ J Bioorg Chem</addtitle><description>This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for
Escherichia coli
Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parameters of interaction of these antibody fragments with cortisol-BSA complex were measured via the biolayer interferometry method. An equilibrium dissociation constant of this interaction K
D
is 5.48 × 10
–10
M. The interaction is reversible and it could be competitively inhibited by free cortisol addition. These results could be used in generating of immunoassays for quantitative cortisol determinations. The in vivo biotinylation system under review is universal and suitable for expression of any biotinylated Fab fragments in the
E. coli
system.</description><subject>Biochemistry</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Bioorganic Chemistry</subject><subject>Escherichia coli</subject><subject>Life Sciences</subject><subject>Organic Chemistry</subject><issn>1068-1620</issn><issn>1608-330X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNp9kEtLAzEUhYMoWKs_wN0s3YzmMY_MUopthYILFdwNN6-aMpPUJF3UX2_KuBNc3cM537lwL0K3BN8TwqqHV4IbThqKSYMJxqw7Q7MseckY_jjPOsflKb9EVzHu8Amq-QyJlXY6QLLeFeBUIT8hgEw62O_J9KYQ1ifrjgMkrYqgpR-FdeBSsQSRS8kKr46FCbAddXZhC9bFVEgfko1-uEYXBoaob37nHL0vn94W63LzsnpePG5KyShJZV3xqmubxkhagdBKKS4xAOuqShFVYaFaY4ymDa4ZpaxWQoiWyFq2tZEt4WyO7qa9--C_DjqmfrRR6mEAp_0h9oRz3rWMY5pRMqEy-BiDNv0-2BHCsSe4P_2z__PP3KFTJ2bWbXXod_4QXL7on9IPkEB5Uw</recordid><startdate>2016</startdate><enddate>2016</enddate><creator>Dormeshkin, D. O.</creator><creator>Kuprienko, O. S.</creator><creator>Svirid, A. V.</creator><creator>Gilep, A. A.</creator><creator>Sviridov, O. V.</creator><creator>Usanov, S. A.</creator><general>Pleiades Publishing</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>2016</creationdate><title>Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol</title><author>Dormeshkin, D. O. ; Kuprienko, O. S. ; Svirid, A. V. ; Gilep, A. A. ; Sviridov, O. V. ; Usanov, S. A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c321t-54849766fc24abeddd8c0aa3944d1d40bd7fffe260532235dbbb71c5c75fc7183</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Biochemistry</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Bioorganic Chemistry</topic><topic>Escherichia coli</topic><topic>Life Sciences</topic><topic>Organic Chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dormeshkin, D. O.</creatorcontrib><creatorcontrib>Kuprienko, O. S.</creatorcontrib><creatorcontrib>Svirid, A. V.</creatorcontrib><creatorcontrib>Gilep, A. A.</creatorcontrib><creatorcontrib>Sviridov, O. V.</creatorcontrib><creatorcontrib>Usanov, S. A.</creatorcontrib><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Russian journal of bioorganic chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dormeshkin, D. O.</au><au>Kuprienko, O. S.</au><au>Svirid, A. V.</au><au>Gilep, A. A.</au><au>Sviridov, O. V.</au><au>Usanov, S. A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol</atitle><jtitle>Russian journal of bioorganic chemistry</jtitle><stitle>Russ J Bioorg Chem</stitle><date>2016</date><risdate>2016</risdate><volume>42</volume><issue>1</issue><spage>22</spage><epage>28</epage><pages>22-28</pages><issn>1068-1620</issn><eissn>1608-330X</eissn><abstract>This paper describes the in vivo generation method of biotinylated recombinant Fab antibody fragments. The original molecular vectors for
Escherichia coli
Fab fragments expression were designed. In vivo biotinylated recombinant antibody Fab fragment against cortisol was generated. The kinetic parameters of interaction of these antibody fragments with cortisol-BSA complex were measured via the biolayer interferometry method. An equilibrium dissociation constant of this interaction K
D
is 5.48 × 10
–10
M. The interaction is reversible and it could be competitively inhibited by free cortisol addition. These results could be used in generating of immunoassays for quantitative cortisol determinations. The in vivo biotinylation system under review is universal and suitable for expression of any biotinylated Fab fragments in the
E. coli
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| language | eng |
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| source | Springer Link |
| subjects | Biochemistry Biomedical and Life Sciences Biomedicine Bioorganic Chemistry Escherichia coli Life Sciences Organic Chemistry |
| title | Generation and characterization of biotinylated recombinant Fab antibody fragment against cortisol |
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