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{"title"=>"Plant-made trastuzumab (Herceptin) inhibits HER2/Neu+ cell proliferation and retards tumor growth", "type"=>"journal", "authors"=>[{"first_name"=>"Tatiana V.", "last_name"=>"Komarova", "scopus_author_id"=>"7005623058"}, {"first_name"=>"Vyacheslav S.", "last_name"=>"Kosorukov", "scopus_author_id"=>"6505962801"}, {"first_name"=>"Olga Y.", "last_name"=>"Frolova", "scopus_author_id"=>"7006584649"}, {"first_name"=>"Igor V.", "last_name"=>"Petrunia", "scopus_author_id"=>"24491077100"}, {"first_name"=>"Ksenia A.", "last_name"=>"Skrypnik", "scopus_author_id"=>"37010028500"}, {"first_name"=>"Yuri Y.", "last_name"=>"Gleba", "scopus_author_id"=>"7003670117"}, {"first_name"=>"Yuri L.", "last_name"=>"Dorokhov", "scopus_author_id"=>"6603938889"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-79952299823", "sgr"=>"79952299823", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0017541", "pmid"=>"21390232", "isbn"=>"1932-6203", "pui"=>"361382340"}, "id"=>"f0146bed-856c-34d9-9d74-c50f922b6aa9", "abstract"=>"BACKGROUND: Plant biotechnology provides a valuable contribution to global health, in part because it can decrease the cost of pharmaceutical products. Breast cancer can now be successfully treated by a humanized monoclonal antibody (mAb), trastuzumab (Herceptin). A course of treatment, however, is expensive and requires repeated administrations of the mAb. Here we used an Agrobacterium-mediated transient expression system to produce trastuzumab in plant cells.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: We describe the cloning and expression of gene constructs in Nicotiana benthamiana plants using intron-optimized Tobacco mosaic virus- and Potato virus X-based vectors encoding, respectively, the heavy and light chains of trastuzumab. Full-size antibodies extracted and purified from plant tissues were tested for functionality and specificity by (i) binding to HER2/neu on the surface of a human mammary gland adenocarcinoma cell line, SK-BR-3, in fluorescence-activated cell sorting assay and (ii) testing the in vitro and in vivo inhibition of HER-2-expressing cancer cell proliferation. We show that plant-made trastuzumab (PMT) bound to the Her2/neu oncoprotein of SK-BR-3 cells and efficiently inhibited SK-BR-3 cell proliferation. Furthermore, mouse intraperitoneal PMT administration retarded the growth of xenografted tumors derived from human ovarian cancer SKOV3 Her2+ cells.\\n\\nCONCLUSIONS/SIGNIFICANCE: We conclude that PMT is active in suppression of cell proliferation and tumor growth.", "link"=>"http://www.mendeley.com/research/plantmade-trastuzumab-herceptin-inhibits-her2neu-cell-proliferation-retards-tumor-growth", "reader_count"=>58, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Librarian"=>2, "Student > Doctoral Student"=>1, "Researcher"=>20, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>4, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>9, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Librarian"=>2, "Student > Doctoral Student"=>1, "Researcher"=>20, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>4, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>9, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>2, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>8, "Materials Science"=>1, "Agricultural and Biological Sciences"=>35, "Medicine and Dentistry"=>4, "Design"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>3, "Social Sciences"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Design"=>{"Design"=>1}, "Engineering"=>{"Engineering"=>2}, "Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Chemistry"=>{"Chemistry"=>3}, "Social Sciences"=>{"Social Sciences"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>35}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1}, "group_count"=>5}

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• http://doi.org/10.1016/j.jbiosc.2017.04.007
• http://doi.org/10.2174/0929867325666171212093257
• http://doi.org/10.1111/pbi.12006
• http://doi.org/10.3389/fpls.2017.02137
• http://doi.org/10.1016/j.pmpp.2018.03.004
• http://doi.org/10.1073/pnas.1718172115
• http://doi.org/10.2144/fsoa-2019-0001
• http://doi.org/10.17650/1726-9784-2018-17-1-95-100
• http://doi.org/10.1007/s12257-011-0636-z
• http://doi.org/10.21519/0234-2758-2019-35-2-49-57
• http://doi.org/10.3390/agriculture5040901
• http://doi.org/10.4161/mabs.3.3.15532
• http://doi.org/10.3109/13880209.2015.1048371
• http://doi.org/10.1016/j.jprot.2013.06.009
• http://doi.org/10.1016/j.apradiso.2018.10.021
• http://doi.org/10.17650/1726-9784-2015-14-2-105-112
• http://doi.org/10.1134/S0006297916100102
• http://doi.org/10.1134/S0006297917040137
• http://doi.org/10.1007/s13580-018-0047-6
• http://doi.org/10.3390/vaccines6010011
• http://doi.org/10.1038/s41598-019-52507-9
• http://doi.org/10.1007/s00253-017-8241-5

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/793579"], "description"=>"<p><b>A</b> – Schematic representation of 35S-based light- (LC) and heavy-(HC) chain- expressing vectors 35S-LC and 35S-HC, respectively. 35S – Cauliflower mosaic virus 35S promoter, T – terminator of transcription, RB and LB – right and left borders from Ti-plasmid. <b>B</b> – Coomassie blue-stained SDS-PAGE proteins before purification (lane 1) and eluted fractions (3–11) obtained after protein A affinity chromatography, lane 2 – flow through after the first loading on the protein A column. M, molecular weight markers. <b>C</b>, <b>D</b> - Western blot analysis of PMT under reducing conditions, developed with anti-gamma (<b>C</b>) and -kappa (<b>D</b>)-chain-specific antibodies.</p>", "links"=>[], "tags"=>["assembled", "pmt", "leaves", "co-injected", "35s-based", "light-", "heavy-chain-expressing"], "article_id"=>463934, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g001", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Production_of_assembled_PMT_in_N_benthamiana_leaves_co_injected_with_35S_based_light_and_heavy_chain_expressing_vectors_/463934", "title"=>"Production of assembled PMT in <i>N. benthamiana</i> leaves co-injected with 35S-based light- and heavy-chain-expressing vectors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:05:34"}
• {"files"=>["https://ndownloader.figshare.com/files/793895"], "description"=>"<p>Flow cytometry analysis of SK-BR-3 cells expressing HER2/neu incubated with trastuzumab (A–C) and PMT (D–F) in the following concentrations: 10 µg/ml (A,D), 1 µg/ml (B,E), and 0.1 µg/ml (C,F). Cells incubated only with secondary reagents were included as a control (open peak). Shadowed areas show specific binding. The percentage of cell surface expression of HER2/neu in SK-BR-3 cells is shown. These data represent three separate experiments.</p>", "links"=>[], "tags"=>["pmt", "binding"], "article_id"=>464265, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g004", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Examination_of_PMT_binding_to_HER2_neu_/464265", "title"=>"Examination of PMT binding to HER2/neu.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:11:05"}
• {"files"=>["https://ndownloader.figshare.com/files/793972"], "description"=>"<p>Growth inhibition effect of PMT compared to trastuzumab (Herceptin, Hoffmann-La Roche) and rituximab (Hoffmann-La Roche) as a negative control. This assay was repeated at least three times.</p>", "links"=>[], "tags"=>["pmt", "cancer", "sk-br-3", "mtt"], "article_id"=>464341, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g005", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_PMT_on_growth_of_the_breast_cancer_cell_line_SK_BR_3_in_MTT_assays_/464341", "title"=>"Effects of PMT on growth of the breast cancer cell line SK-BR-3 in MTT assays.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:12:21"}
• {"files"=>["https://ndownloader.figshare.com/files/794035"], "description"=>"<p>The treatment groups received their first doses (20 mg/kg) of PMT (n = 7) and trastuzumab (n = 10) in saline solution i.p. 6 days after SKOV3 implantation, and then for 16 days, they received 8 consecutive injections (10 mg/kg). The control group (n = 34) received saline solution. Tumor volumes were recorded in intervals 10–14, 18–22 and 23–27 days after SKOV3 implantation using a caliper. Data are the mean ± standard deviations from two independent experiments. Asterisk shows P<0.05 by the unpaired two tailed Student's t-test for statistical significance of difference between the PMT and trastuzumab treatment and control.</p>", "links"=>[], "tags"=>["inhibits", "xenograft", "ovarian"], "article_id"=>464399, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g006", "stats"=>{"downloads"=>2, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PMT_inhibits_tumor_growth_in_a_xenograft_model_of_human_Her2_ovarian_cancer_/464399", "title"=>"PMT inhibits tumor growth in a xenograft model of human Her2+ ovarian cancer.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:13:19"}
• {"files"=>["https://ndownloader.figshare.com/files/397149", "https://ndownloader.figshare.com/files/397184"], "description"=>"<div><h3>Background</h3><p>Plant biotechnology provides a valuable contribution to global health, in part because it can decrease the cost of pharmaceutical products. Breast cancer can now be successfully treated by a humanized monoclonal antibody (mAb), trastuzumab (Herceptin). A course of treatment, however, is expensive and requires repeated administrations of the mAb. Here we used an <em>Agrobacterium</em>-mediated transient expression system to produce trastuzumab in plant cells.</p> <h3>Methodology/Principal Findings</h3><p>We describe the cloning and expression of gene constructs in <em>Nicotiana benthamiana</em> plants using intron-optimized <em>Tobacco mosaic virus</em>- and <em>Potato virus X</em>-based vectors encoding, respectively, the heavy and light chains of trastuzumab. Full-size antibodies extracted and purified from plant tissues were tested for functionality and specificity by (i) binding to HER2/neu on the surface of a human mammary gland adenocarcinoma cell line, SK-BR-3, in fluorescence-activated cell sorting assay and (ii) testing the <em>in vitro</em> and <em>in vivo</em> inhibition of HER-2-expressing cancer cell proliferation. We show that plant-made trastuzumab (PMT) bound to the Her2/neu oncoprotein of SK-BR-3 cells and efficiently inhibited SK-BR-3 cell proliferation. Furthermore, mouse intraperitoneal PMT administration retarded the growth of xenografted tumors derived from human ovarian cancer SKOV3 Her2+ cells.</p> <h3>Conclusions/Significance</h3><p>We conclude that PMT is active in suppression of cell proliferation and tumor growth.</p> </div>", "links"=>[], "tags"=>["plant-made", "trastuzumab", "inhibits", "proliferation", "retards"], "article_id"=>138446, "categories"=>["Cancer", "Biotechnology", "Cell Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0017541.s001", "https://dx.doi.org/10.1371/journal.pone.0017541.s002"], "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Plant_Made_Trastuzumab_Herceptin_Inhibits_HER2_Neu_Cell_Proliferation_and_Retards_Tumor_Growth/138446", "title"=>"Plant-Made Trastuzumab (Herceptin) Inhibits HER2/Neu+ Cell Proliferation and Retards Tumor Growth", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-03-03 02:20:46"}
• {"files"=>["https://ndownloader.figshare.com/files/793836"], "description"=>"<p>Comparative binding of trastuzumab and PMT to the HER2/neu-specific cyclic synthetic peptide 563CYC (CHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVA) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0017541#pone.0017541-Garrett1\" target=\"_blank\">[30]</a>. Microtiter wells were coated overnight with 2 µg/ml peptide and then blocked with 1% BSA for 1 h. The mAbs were then added to plates at a concentration of 250 µg/ml and serially diluted 1∶1 with phosphate buffered saline (PBS). Bound mAb was detected with HRP-conjugated anti-human IgG and then with substrate.</p>", "links"=>[], "tags"=>["pmt", "peptide"], "article_id"=>464203, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g003", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Binding_of_PMT_to_a_HER2_neu_peptide_mimotope_/464203", "title"=>"Binding of PMT to a HER2/neu peptide mimotope.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:10:03"}
• {"files"=>["https://ndownloader.figshare.com/files/793726"], "description"=>"<p><b>A</b> – Schematic representation of PVX- and crTMV-based vectors. LB and RB, binary vector left and right borders, respectively; 35S, 35S promoter; Act 2, <i>Arabidopsis</i> actin 2 promoter; T, <i>nos</i> terminator; RdRp, RNA-dependent RNA polymerase; Bars 1–8, introns; MP, TMV movement protein; 25K, 12K, 8K, PVX movement protein genes. <b>B–E</b> - Western blot analysis of purified PMT. Purification of mAbs on protein A sepharose. Proteins were separated in a 10% polyacrylamide gel under non-reducing conditions (<b>B</b>, <b>C</b>) and in a 12% gel under reducing conditions (<b>D</b>, <b>E</b>) and transferred to a PVDF membrane. Western blots: <b>B</b> and <b>D</b> were probed with gamma-chain-specific antibodies; membranes <b>C</b> and <b>E</b> were incubated with kappa-chain-specific antibodies. 1–2, fractions from the protein A sepharose column; M, protein molecular weight markers; S, standard - 20 ng hIgG. <b>F</b> - Capillary electrophoresis analysis of PMT in reducing conditions on Agilent 2100 Bioanalyzer. Peak 12 corresponds to HC; peak 8 corresponds to LC. <b>G</b>, <b>H</b> – Comparison of PMT and trastuzumab. Proteins were separated in a 7.5% polyacrylamide gel under non-reducing conditions (<b>G</b>) and in a 12% gel under reducing conditions (<b>H</b>) and stained with Coomassie blue. <b>I</b> - RP-HPLC trace analysis of PMT and trastuzumab. The linear gradient was 0–60% acetonitrile for 20 min and then 60–100% acetonitrile for 5 min; the flow rate was 80 µL·min⁻¹. The buffer blank was 10 mM Na-phosphate (pH 7.0). Absorbance at 214 nm and 280 nm is shown.</p>", "links"=>[], "tags"=>["purification", "assembled", "pmt", "leaves", "co-injected", "light-chain-encoding", "pvx-based", "heavy-chain-encoding", "tmv-based"], "article_id"=>464088, "categories"=>["Cancer", "Biotechnology", "Plant Biology"], "users"=>["Tatiana V. Komarova", "Vyacheslav S. Kosorukov", "Olga Y. Frolova", "Igor V. Petrunia", "Ksenia A. Skrypnik", "Yuri Y. Gleba", "Yuri L. Dorokhov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0017541.g002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Accumulation_and_purification_of_assembled_PMT_in_N_benthamiana_leaves_co_injected_with_light_chain_encoding_PVX_based_and_heavy_chain_encoding_TMV_based_vectors_/464088", "title"=>"Accumulation and purification of assembled PMT in <i>N. benthamiana</i> leaves co-injected with light-chain-encoding PVX-based and heavy-chain-encoding TMV-based vectors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-03-03 01:08:08"}

PMC Usage Stats | Further Information

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F1000Prime