Detection of Rubisco and mycotoxins as potential contaminants of a plantibody against the hepatitis B surface antigen purified from tobacco
Antibodies have been one of the proteins widely expressed in tobacco plants for pharmaceutical purposes, which demand contaminant free preparations. Rubisco constitutes 40–60% of tobacco leaf soluble proteins; therefore it is the major potential protein contaminant of plantibodies, while mycotoxins...
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| Veröffentlicht in: | Biologicals 2007-10, Vol.35 (4), p.309-315 |
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| creator | Geada, Déborah Valdés, Rodolfo Escobar, Arturo Ares, Dulce M. Torres, Edel Blanco, Reinaldo Ferro, Williams Dorta, Dayamí González, Marcos Alemán, María R. Padilla, Sigifredo Gómez, Leonardo del Castillo, Norma Mendoza, Otto Urquiza, Dioslaida Soria, Yordanka Brito, José Leyva, Alberto Borroto, Carlos Gavilondo, Jorge V. |
| description | Antibodies have been one of the proteins widely expressed in tobacco plants for pharmaceutical purposes, which demand contaminant free preparations. Rubisco constitutes 40–60% of tobacco leaf soluble proteins; therefore it is the major potential protein contaminant of plantibodies, while mycotoxins are toxic compounds that could be introduced during the biomass production and post-harvest stages with important consequences to human health. The objective of this paper was to investigate whether Rubisco and mycotoxins are present in Plantibody HB-01 preparations used in the immunopurification of the hepatitis B surface antigen. Rubisco was purified from
Nicotiana tabacum yielding 154
μg of protein per gram of leaves and purity over 95%. Among mouse monoclonal antibodies generated against this enzyme, the CBSS.Rub-2 was selected for its immunodetection. It recognizes a conserved sequential epitope of Rubisco large subunit with an affinity constant of 0.13
×
10
8
M
−1. Rubisco quantification limit was 1
μg spreading to the measurement of this contaminant less than 4% of plantibodies samples. Additionally, according to a Reverse Phase-HPLC used to measure the level of adventitiously introduced contaminants, it can be concluded that aflatoxins B1, B2, G1 and G2 were undetected in the purified Plantibody HB-01 samples. |
| doi_str_mv | 10.1016/j.biologicals.2007.02.007 |
| format | Article |
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Nicotiana tabacum yielding 154
μg of protein per gram of leaves and purity over 95%. Among mouse monoclonal antibodies generated against this enzyme, the CBSS.Rub-2 was selected for its immunodetection. It recognizes a conserved sequential epitope of Rubisco large subunit with an affinity constant of 0.13
×
10
8
M
−1. Rubisco quantification limit was 1
μg spreading to the measurement of this contaminant less than 4% of plantibodies samples. Additionally, according to a Reverse Phase-HPLC used to measure the level of adventitiously introduced contaminants, it can be concluded that aflatoxins B1, B2, G1 and G2 were undetected in the purified Plantibody HB-01 samples.</description><identifier>ISSN: 1045-1056</identifier><identifier>EISSN: 1095-8320</identifier><identifier>DOI: 10.1016/j.biologicals.2007.02.007</identifier><identifier>PMID: 17644409</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Aflatoxins ; Aflatoxins - analysis ; Aflatoxins - chemistry ; Animals ; Antibodies, Monoclonal - genetics ; Antibodies, Monoclonal - isolation & purification ; Blotting, Western ; Chromatography, High Pressure Liquid - methods ; Chromatography, High Pressure Liquid - statistics & numerical data ; Drug Contamination ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Antibodies - genetics ; Hepatitis B Antibodies - isolation & purification ; Hepatitis B Surface Antigens - immunology ; Humans ; Mice ; Monoclonal antibody ; Nicotiana - genetics ; Nicotiana - immunology ; Nicotiana tabacum ; Plantibodies - genetics ; Plantibodies - isolation & purification ; Plantibody ; Plants, Genetically Modified ; Ribulose-Bisphosphate Carboxylase - analysis ; Rubisco</subject><ispartof>Biologicals, 2007-10, Vol.35 (4), p.309-315</ispartof><rights>2007 The International Association for Biologicals</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c406t-8af82d9f5a1cb77ff8d515aaec981e0421d27304d8104bb0243577e901fead483</citedby><cites>FETCH-LOGICAL-c406t-8af82d9f5a1cb77ff8d515aaec981e0421d27304d8104bb0243577e901fead483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.biologicals.2007.02.007$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17644409$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Geada, Déborah</creatorcontrib><creatorcontrib>Valdés, Rodolfo</creatorcontrib><creatorcontrib>Escobar, Arturo</creatorcontrib><creatorcontrib>Ares, Dulce M.</creatorcontrib><creatorcontrib>Torres, Edel</creatorcontrib><creatorcontrib>Blanco, Reinaldo</creatorcontrib><creatorcontrib>Ferro, Williams</creatorcontrib><creatorcontrib>Dorta, Dayamí</creatorcontrib><creatorcontrib>González, Marcos</creatorcontrib><creatorcontrib>Alemán, María R.</creatorcontrib><creatorcontrib>Padilla, Sigifredo</creatorcontrib><creatorcontrib>Gómez, Leonardo</creatorcontrib><creatorcontrib>del Castillo, Norma</creatorcontrib><creatorcontrib>Mendoza, Otto</creatorcontrib><creatorcontrib>Urquiza, Dioslaida</creatorcontrib><creatorcontrib>Soria, Yordanka</creatorcontrib><creatorcontrib>Brito, José</creatorcontrib><creatorcontrib>Leyva, Alberto</creatorcontrib><creatorcontrib>Borroto, Carlos</creatorcontrib><creatorcontrib>Gavilondo, Jorge V.</creatorcontrib><title>Detection of Rubisco and mycotoxins as potential contaminants of a plantibody against the hepatitis B surface antigen purified from tobacco</title><title>Biologicals</title><addtitle>Biologicals</addtitle><description>Antibodies have been one of the proteins widely expressed in tobacco plants for pharmaceutical purposes, which demand contaminant free preparations. Rubisco constitutes 40–60% of tobacco leaf soluble proteins; therefore it is the major potential protein contaminant of plantibodies, while mycotoxins are toxic compounds that could be introduced during the biomass production and post-harvest stages with important consequences to human health. The objective of this paper was to investigate whether Rubisco and mycotoxins are present in Plantibody HB-01 preparations used in the immunopurification of the hepatitis B surface antigen. Rubisco was purified from
Nicotiana tabacum yielding 154
μg of protein per gram of leaves and purity over 95%. Among mouse monoclonal antibodies generated against this enzyme, the CBSS.Rub-2 was selected for its immunodetection. It recognizes a conserved sequential epitope of Rubisco large subunit with an affinity constant of 0.13
×
10
8
M
−1. Rubisco quantification limit was 1
μg spreading to the measurement of this contaminant less than 4% of plantibodies samples. Additionally, according to a Reverse Phase-HPLC used to measure the level of adventitiously introduced contaminants, it can be concluded that aflatoxins B1, B2, G1 and G2 were undetected in the purified Plantibody HB-01 samples.</description><subject>Aflatoxins</subject><subject>Aflatoxins - analysis</subject><subject>Aflatoxins - chemistry</subject><subject>Animals</subject><subject>Antibodies, Monoclonal - genetics</subject><subject>Antibodies, Monoclonal - isolation & purification</subject><subject>Blotting, Western</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Chromatography, High Pressure Liquid - statistics & numerical data</subject><subject>Drug Contamination</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Hepatitis B Antibodies - genetics</subject><subject>Hepatitis B Antibodies - isolation & purification</subject><subject>Hepatitis B Surface Antigens - immunology</subject><subject>Humans</subject><subject>Mice</subject><subject>Monoclonal antibody</subject><subject>Nicotiana - genetics</subject><subject>Nicotiana - immunology</subject><subject>Nicotiana tabacum</subject><subject>Plantibodies - genetics</subject><subject>Plantibodies - isolation & purification</subject><subject>Plantibody</subject><subject>Plants, Genetically Modified</subject><subject>Ribulose-Bisphosphate Carboxylase - analysis</subject><subject>Rubisco</subject><issn>1045-1056</issn><issn>1095-8320</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc2O1DAQhC0EYn_gFZC5cEtoO84kOcIssEgrISE4W47dnvUosYPtIOYZeGkczUhw5FR9-Kpb1UXIawY1A7Z7e6xHF6ZwcFpNqeYAXQ28LvKEXDMY2qpvODzdZtFWDNrdFblJ6QjAmOjEc3LFup0QAoZr8vsOM-rsgqfB0q_r6JIOVHlD55MOOfxyPlGV6BIy-uzURHXwWc3OK5_T5lF0mcrsxmBOVB1UMWSaH5E-4qKyyy7R9zSt0SqNdAMP6OmyRmcdGmpjmGkOo9I6vCDPbAmELy96S75__PBtf189fPn0ef_uodICdrnqle25GWyrmB67ztretKxVCvXQMwTBmeFdA8L0Jf84AhdN23U4ALOojOibW_LmvHeJ4ceKKcu5pMapxMCwJsmBtzA0vIDDGdQxpBTRyiW6WcWTZCC3JuRR_tOE3JqQwGWR4n11ObKOM5q_zsvrC7A_A1ii_nQYZdIOvUbjYmlEmuD-48wfZ5OjVg</recordid><startdate>20071001</startdate><enddate>20071001</enddate><creator>Geada, Déborah</creator><creator>Valdés, Rodolfo</creator><creator>Escobar, Arturo</creator><creator>Ares, Dulce M.</creator><creator>Torres, Edel</creator><creator>Blanco, Reinaldo</creator><creator>Ferro, Williams</creator><creator>Dorta, Dayamí</creator><creator>González, Marcos</creator><creator>Alemán, María R.</creator><creator>Padilla, Sigifredo</creator><creator>Gómez, Leonardo</creator><creator>del Castillo, Norma</creator><creator>Mendoza, Otto</creator><creator>Urquiza, Dioslaida</creator><creator>Soria, Yordanka</creator><creator>Brito, José</creator><creator>Leyva, Alberto</creator><creator>Borroto, Carlos</creator><creator>Gavilondo, Jorge V.</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7T7</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope></search><sort><creationdate>20071001</creationdate><title>Detection of Rubisco and mycotoxins as potential contaminants of a plantibody against the hepatitis B surface antigen purified from tobacco</title><author>Geada, Déborah ; Valdés, Rodolfo ; Escobar, Arturo ; Ares, Dulce M. ; Torres, Edel ; Blanco, Reinaldo ; Ferro, Williams ; Dorta, Dayamí ; González, Marcos ; Alemán, María R. ; Padilla, Sigifredo ; Gómez, Leonardo ; del Castillo, Norma ; Mendoza, Otto ; Urquiza, Dioslaida ; Soria, Yordanka ; Brito, José ; Leyva, Alberto ; Borroto, Carlos ; Gavilondo, Jorge V.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c406t-8af82d9f5a1cb77ff8d515aaec981e0421d27304d8104bb0243577e901fead483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Aflatoxins</topic><topic>Aflatoxins - analysis</topic><topic>Aflatoxins - chemistry</topic><topic>Animals</topic><topic>Antibodies, Monoclonal - genetics</topic><topic>Antibodies, Monoclonal - isolation & purification</topic><topic>Blotting, Western</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Chromatography, High Pressure Liquid - statistics & numerical data</topic><topic>Drug Contamination</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Hepatitis B Antibodies - genetics</topic><topic>Hepatitis B Antibodies - isolation & purification</topic><topic>Hepatitis B Surface Antigens - immunology</topic><topic>Humans</topic><topic>Mice</topic><topic>Monoclonal antibody</topic><topic>Nicotiana - genetics</topic><topic>Nicotiana - immunology</topic><topic>Nicotiana tabacum</topic><topic>Plantibodies - genetics</topic><topic>Plantibodies - isolation & purification</topic><topic>Plantibody</topic><topic>Plants, Genetically Modified</topic><topic>Ribulose-Bisphosphate Carboxylase - analysis</topic><topic>Rubisco</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Geada, Déborah</creatorcontrib><creatorcontrib>Valdés, Rodolfo</creatorcontrib><creatorcontrib>Escobar, Arturo</creatorcontrib><creatorcontrib>Ares, Dulce M.</creatorcontrib><creatorcontrib>Torres, Edel</creatorcontrib><creatorcontrib>Blanco, Reinaldo</creatorcontrib><creatorcontrib>Ferro, Williams</creatorcontrib><creatorcontrib>Dorta, Dayamí</creatorcontrib><creatorcontrib>González, Marcos</creatorcontrib><creatorcontrib>Alemán, María R.</creatorcontrib><creatorcontrib>Padilla, Sigifredo</creatorcontrib><creatorcontrib>Gómez, Leonardo</creatorcontrib><creatorcontrib>del Castillo, Norma</creatorcontrib><creatorcontrib>Mendoza, Otto</creatorcontrib><creatorcontrib>Urquiza, Dioslaida</creatorcontrib><creatorcontrib>Soria, Yordanka</creatorcontrib><creatorcontrib>Brito, José</creatorcontrib><creatorcontrib>Leyva, Alberto</creatorcontrib><creatorcontrib>Borroto, Carlos</creatorcontrib><creatorcontrib>Gavilondo, Jorge V.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Biologicals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Geada, Déborah</au><au>Valdés, Rodolfo</au><au>Escobar, Arturo</au><au>Ares, Dulce M.</au><au>Torres, Edel</au><au>Blanco, Reinaldo</au><au>Ferro, Williams</au><au>Dorta, Dayamí</au><au>González, Marcos</au><au>Alemán, María R.</au><au>Padilla, Sigifredo</au><au>Gómez, Leonardo</au><au>del Castillo, Norma</au><au>Mendoza, Otto</au><au>Urquiza, Dioslaida</au><au>Soria, Yordanka</au><au>Brito, José</au><au>Leyva, Alberto</au><au>Borroto, Carlos</au><au>Gavilondo, Jorge V.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Rubisco and mycotoxins as potential contaminants of a plantibody against the hepatitis B surface antigen purified from tobacco</atitle><jtitle>Biologicals</jtitle><addtitle>Biologicals</addtitle><date>2007-10-01</date><risdate>2007</risdate><volume>35</volume><issue>4</issue><spage>309</spage><epage>315</epage><pages>309-315</pages><issn>1045-1056</issn><eissn>1095-8320</eissn><abstract>Antibodies have been one of the proteins widely expressed in tobacco plants for pharmaceutical purposes, which demand contaminant free preparations. Rubisco constitutes 40–60% of tobacco leaf soluble proteins; therefore it is the major potential protein contaminant of plantibodies, while mycotoxins are toxic compounds that could be introduced during the biomass production and post-harvest stages with important consequences to human health. The objective of this paper was to investigate whether Rubisco and mycotoxins are present in Plantibody HB-01 preparations used in the immunopurification of the hepatitis B surface antigen. Rubisco was purified from
Nicotiana tabacum yielding 154
μg of protein per gram of leaves and purity over 95%. Among mouse monoclonal antibodies generated against this enzyme, the CBSS.Rub-2 was selected for its immunodetection. It recognizes a conserved sequential epitope of Rubisco large subunit with an affinity constant of 0.13
×
10
8
M
−1. Rubisco quantification limit was 1
μg spreading to the measurement of this contaminant less than 4% of plantibodies samples. Additionally, according to a Reverse Phase-HPLC used to measure the level of adventitiously introduced contaminants, it can be concluded that aflatoxins B1, B2, G1 and G2 were undetected in the purified Plantibody HB-01 samples.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>17644409</pmid><doi>10.1016/j.biologicals.2007.02.007</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Biologicals, 2007-10, Vol.35 (4), p.309-315 |
| issn | 1045-1056 1095-8320 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Aflatoxins Aflatoxins - analysis Aflatoxins - chemistry Animals Antibodies, Monoclonal - genetics Antibodies, Monoclonal - isolation & purification Blotting, Western Chromatography, High Pressure Liquid - methods Chromatography, High Pressure Liquid - statistics & numerical data Drug Contamination Enzyme-Linked Immunosorbent Assay Hepatitis B Antibodies - genetics Hepatitis B Antibodies - isolation & purification Hepatitis B Surface Antigens - immunology Humans Mice Monoclonal antibody Nicotiana - genetics Nicotiana - immunology Nicotiana tabacum Plantibodies - genetics Plantibodies - isolation & purification Plantibody Plants, Genetically Modified Ribulose-Bisphosphate Carboxylase - analysis Rubisco |
| title | Detection of Rubisco and mycotoxins as potential contaminants of a plantibody against the hepatitis B surface antigen purified from tobacco |
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