Recombinant Production of Human Interleukin 6 in Escherichia coli
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{"title"=>"Recombinant Production of Human Interleukin 6 in Escherichia coli", "type"=>"journal", "authors"=>[{"first_name"=>"Henrik", "last_name"=>"Nausch", "scopus_author_id"=>"6507003753"}, {"first_name"=>"Jana", "last_name"=>"Huckauf", "scopus_author_id"=>"6507489015"}, {"first_name"=>"Roswitha", "last_name"=>"Koslowski", "scopus_author_id"=>"55481161100"}, {"first_name"=>"Udo", "last_name"=>"Meyer", "scopus_author_id"=>"7202256661"}, {"first_name"=>"Inge", "last_name"=>"Broer", "scopus_author_id"=>"6602556049"}, {"first_name"=>"Heike", "last_name"=>"Mikschofsky", "scopus_author_id"=>"6506430748"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84872824121", "doi"=>"10.1371/journal.pone.0054933", "pui"=>"368176496", "issn"=>"19326203", "pmid"=>"23372793", "isbn"=>"1932-6203", "scopus"=>"2-s2.0-84872824121"}, "id"=>"93315c3f-c6e4-3a92-80eb-936065cc835c", "abstract"=>"In this study, we compared basic expression approaches for the efficient expression of bioactive recombinant human interleukin-6 (IL6), as an example for a difficult-to-express protein. We tested these approaches in a laboratory scale in order to pioneer the commercial production of this protein in Escherichia coli (E. coli). Among the various strategies, which were tested under Research and Development (R&D) conditions, aggregation-prone IL6 was solubilized most effectively by co-expressing cytoplasmic chaperones. Expression of a Glutathion-S-Transferase (GST) fusion protein was not efficient to increase IL6 solubility. Alteration of the cultivation temperature significantly increased the solubility in both cases, whereas reduced concentrations of IPTG to induce expression of the T7lac-promotor only had a positive effect on chaperone-assisted expression. The biological activity was comparable to that of commercial IL6. Targeting the expressed protein to an oxidizing environment was not effective in the generation of soluble IL6. Taken together, the presence of chaperones and a lowered cultivation temperature seem effective to isolate large quantities of soluble IL6. This approach led to in vivo soluble, functional protein fractions and reduces purification and refolding requirements caused by downstream purification procedures. The final yield of soluble recombinant protein averaged approximately 2.6 mg IL6/liter of cell culture. These findings might be beneficial for the development of the large-scale production of IL6 under the conditions of current good manufacturing practice (cGMP).", "link"=>"http://www.mendeley.com/research/recombinant-production-human-interleukin-6-escherichia-coli-1", "reader_count"=>38, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>10, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>10, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>13, "Agricultural and Biological Sciences"=>18, "Medicine and Dentistry"=>2, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>1, "Social Sciences"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>13}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Portugal"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

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  • {"files"=>["https://ndownloader.figshare.com/files/503300"], "description"=>"<p>(S) soluble and (I) insoluble fraction of IL6.</p>", "links"=>[], "tags"=>["blotting", "il6-expressing", "strains", "bl21", "origami"], "article_id"=>173813, "categories"=>["Physiology", "Biotechnology", "Biochemistry", "Immunology"], "users"=>["Henrik Nausch", "Jana Huckauf", "Roswitha Koslowski", "Udo Meyer", "Inge Broer", "Heike Mikschofsky"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0054933.g002", "stats"=>{"downloads"=>2, "page_views"=>29, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blotting_analysis_of_the_IL6_expressing_E_coli_strains_BL21_and_Origami_2_/173813", "title"=>"Western blotting analysis of the IL6-expressing <i>E. coli</i> strains BL21 and Origami 2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:41:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/503707"], "description"=>"<p>Recombinant strains used in this work.</p>", "links"=>[], "tags"=>["strains"], "article_id"=>174230, "categories"=>["Physiology", "Biotechnology", "Biochemistry", "Immunology"], "users"=>["Henrik Nausch", "Jana Huckauf", "Roswitha Koslowski", "Udo Meyer", "Inge Broer", "Heike Mikschofsky"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0054933.t001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Recombinant_strains_used_in_this_work_/174230", "title"=>"Recombinant strains used in this work.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-19 16:43:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/503433"], "description"=>"<p>(S) soluble and (I) insoluble fraction of IL6, expressed with and without the concomitant overexpression of endogenous cytoplasmic chaperones DnaK, DnaJ, GrpE, GroES, GroEL (C-Chaperones) or periplasmic chaperones DsbA, DsbC, SurA, FkpA (P-Chaperones) at 37°C and 22°C, respectively. Additionally, IL6 was expressed fused to GST in the cytoplasm.</p>", "links"=>[], "tags"=>["blotting", "il6", "cytoplasm", "periplasm"], "article_id"=>173945, "categories"=>["Physiology", "Biotechnology", "Biochemistry", "Immunology"], "users"=>["Henrik Nausch", "Jana Huckauf", "Roswitha Koslowski", "Udo Meyer", "Inge Broer", "Heike Mikschofsky"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0054933.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blotting_analysis_of_the_IL6_expression_in_the_cytoplasm_and_periplasm_of_E_coli_BL21_/173945", "title"=>"Western blotting analysis of the IL6 expression in the cytoplasm and periplasm of <i>E. coli</i> BL21.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:42:06"}
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  • {"unique-ip"=>"26", "full-text"=>"29", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"26", "full-text"=>"26", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"42", "full-text"=>"56", "pdf"=>"13", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"43", "full-text"=>"60", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"22", "full-text"=>"20", "pdf"=>"10", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"51", "full-text"=>"58", "pdf"=>"7", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"30", "full-text"=>"34", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"26", "full-text"=>"29", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}

Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[269, 466, 588, 697, 800, 896, 988, 1076, 1165, 1254, 1340, 1417]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[272, 472, 600, 713, 815, 911, 1004, 1094, 1185, 1273, 1358, 1441]}, {"subject_area"=>"/Biology and life sciences/Microbiology", "average_usage"=>[293, 503, 638, 755, 861, 960, 1056, 1146, 1239, 1323, 1403, 1491, 1568]}, {"subject_area"=>"/Physical sciences", "average_usage"=>[254, 431, 547, 651, 748, 842, 932, 1017, 1098, 1178, 1259, 1336, 1404]}, {"subject_area"=>"/Physical sciences/Materials science", "average_usage"=>[250, 417, 535, 649, 758, 857, 952, 1036, 1113, 1206, 1287, 1353, 1429]}]}
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