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All Journal HAYATI Journal of Biosciences Jurnal Veteriner Microbiology Indonesia Pharmaciana: Jurnal Kefarmasian Jurnal Matematika & Sains Journal of Mathematical and Fundamental Sciences Jurnal Sains dan Teknologi Farmasi Jurnal Media Gizi Indonesia (MGI) INDONESIAN JOURNAL OF PHARMACY Jurnal Komputer Terapan ANNALES BOGORIENSES Jurnal Sisfokom (Sistem Informasi dan Komputer) FITOFARMAKA: Jurnal Ilmiah Farmasi Jurnal Respirasi (JR) Darmabakti Cendekia: Journal of Community Service and Engagements Jurnal Layanan Masyarakat (Journal of Public Service) Jurnal Teknosains Kodepena (Kodepena Journal of Technoscience) Narra J Jurnal Teknosains Kodepena (Kodepena Journal of Technoscience) Jurnal Puruhita MEDICINUS Journal of Community Engagement in Health Annales Bogorienses
HENI RACHMAWATI
School of Pharmacy, Institute of Technology Bandung. Jl. Ganesha, 10, Bandung 40132
Agriculture, Biological Sciences & Forestry Humanities Astronomy Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Dentistry Earth & Planetary Sciences Economics, Econometrics & Finance Education Electrical & Electronics Engineering Energy Engineering Environmental Science Health Professions Immunology & microbiology Mathematics Medicine & Pharmacology Neuroscience Nursing Physics Public Health Social Sciences Veterinary Other

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Journal : Microbiology Indonesia

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Optimization of Human Interferon α2b Soluble Protein Overproduction and Primary Recovery of Its Inclusion Bodies RATIH ASMANA NINGRUM; DEBBIE SOFIE RETNONINGRUM; YEYET CAHYATI; HENI RACHMAWATI
Microbiology Indonesia Vol. 5 No. 1 (2011): March 2011
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (141.802 KB) | DOI: 10.5454/mi.5.1.5

Abstract

The hIFN2b open reading frame has been constructed and overexpressed in Escherichia coli BL21(DE3). The yields of protein purified using nickel column from inclusion bodies (IB) and total soluble proteins were 3.46 mg and 2.57 mg in 1 L culture, respectively. This research was aimed to obtain optimal condition for high level overproduction of soluble hIFNα2b as well as primary recovery of hIFN2b from IB. We used two different conditions for obtaining soluble protein, i.e. induction temperatures and inducer concentrations, and three different conditions for inclusion bodies, i.e. centrifugation speeds, washing and solubilizing buffers. Induction using 0.5 mM of isopropyl thiogalactopyranoside at 25 °C yielded 8.9 mg hIFN2b in 1 L culture. The best recovery of IB was achieved when 10 000 g was applied for centrifugation, 1% Triton X-100 in 50 mM Tris Cl pH 8.0 as washing buffer, and 8M guanidine HCl in 50 mM Tris Cl pH 8.0 containing 800 mM 2-mercaptoethanol as solubilizing buffer were used. At this optimal condition the yield of hIFN2b from IB was 28.85 mg in 1 L culture. The total recovery of hIFNα2b at optimal condition was 50% from IB and 14% from soluble protein. hIFN2b from IB was refolded by 9 d dialysis in refolding buffer (0.2 mM EDTA, 0.25 mM ditiothreitol, 50 mM Tris and 0.4 M urea pH 8.0).
Co-Authors Abdillah, Oktaviardi Bityasmawan Agustiyanti, Dian Fitria Agustiyanti, Dian Fitria Alfan Danny Arbianto Ali Iqbal Tawakal Ali Iqbal Tawakal Amalia, Riezki Ambarwati, Rini Amirah Adlia Amirah Adlia Amirah Adlia Anna Smdyah Putri Anna Surgean Veterini Annis Catur Adi Arbianto, Alfan Danny Arbianto, Alfan Danny Arini Setiawati Armay, Edmond Febrinicko Asrul Muhamad Fuad Asrul Muhamad Fuad, Asrul Muhamad Atsarina Anindya Bambang Wahjuprajitno Citra Ariani Edityaningrum, Citra Damar Rasti Adhika Deandra Ardya Regitasari Deandra Ardya Regitasari DEBBIE SOFIE RETNONINGRUM DEBBIE SOFIE RETNONINGRUM Debbie Sofie Retnoningrum Dewi Esti Restiani Dian Fitria Agustiyanti Dwiyatna, Archie Arman Emyr Reisha Ishaura Estherina Juliana Marbun, Estherina Juliana Farapti Farapti Fathrizqita Aghnia Raudhany Fatona Suraya, Fatona Ferry Iskandar Fibriani, Azzania Hamzah Hamzah Herdiani Sulistyo Putri Hidayat, Erzi I Ketut Adnyana Irda Fidrianny Isaura, Emyr Reisha Iskandar, Fery Jessie S. Pamudji Kusnandar Anggadiredja Leonny Yulita Hartiadi Lili Fitriani Magdalena Yuni Kristanti Maharani, Dita Galuh Mahmud Aditya Rifqi Marlina Indriastuti Meirawan, Rizky Fajar Muhammad mahrus zain Nelly Setiawaty Nuthathai Sutthiwong Permatasari, Fitri A. Permatasari, Fitri Aulia Pricillia Sinambela Putri, Dea Mayang Yulia Rachmad Suhanda Rachmadani, Nisa Amanda Rachmat Mauludin Rahmah, Ainur Rahmana E. Kartasasmita Rahmana E. Kartasasmita Rasyidi, Mohammad Fahmi RATIH ASMANA NINGRUM Rehatta, Nancy Margarita Resmi, Juniar Kalpika Restiani, Dewi Esti Restiani, Dewi Esti Rika Hartati Rini Ambarwati S.Pd. M Kes I Ketut Sudiana . Sabar Pambudi Safira Prisya Dewi Sakinah Aljuffrie Salisa, Wizara Santika, Arum Sinda Sari, Ririn Andika Sarmanu, Sarmanu Satuman Satuman Shafiqah Adam Siti Fatimah Zahro Soetomo, Meilisa Keizia Sophi Damayanti Subijanto Marto Soedarmo Subijanto Marto Soedarmo Sukmadjaja Asyarie Sukmadjaja Asyarie Sukmadjaja Asyarie Sulistiawan, Soni Sunarso Syefrida Yulina, Syefrida Taharuddin, Audrey Angelina Putri Tarwadi Tarwadi Tarwadi, Tarwadi Tarwadi, Tarwadi Tjandrawati Mozef Tri Suciati Vidya Anggarini Rahmasari Vidya Anggarini Rahmasari Wenda Novayani Wibowo, Indra Widijiati Widijiati Widjiati Widjiati Widodo Jatim P Widodo Jatim Pudjirahardjo Widodo, Ferri Wijayanti, Ni PAD. Wizara Salisa Wizara Salisa Wizara Salisa Yeyet C. Sumirtapura YEYET CAHYATI Yogie Handoko