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All Journal HAYATI Journal of Biosciences Jurnal Veteriner Microbiology Indonesia Pharmaciana: Jurnal Kefarmasian Medical Journal of Indonesia 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) Narra J 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 Engineering Environmental Science Health Professions Immunology & microbiology Mathematics Medicine & Pharmacology Neuroscience Nursing Physics Public Health Social Sciences Veterinary Other

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Journal : Annales Bogorienses

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DNA Condensation Study of Fully Synthesized Lipopeptide-Based Transfection Agent for Gene Delivery Vehicle Tarwadi, Tarwadi; Rachmawati, Heni; Kartasasmita, Rahmana E.; Pambudi, Sabar; Arbianto, Alfan Danny; Restiani, Dewi Esti; Asyarie, Sukmadjaja
Annales Bogorienses Vol. 22 No. 2 (2018): Annales Bogorienses
Publisher : BRIN

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Abstract

The main requirement of transfection agent has to condense DNA in nanoparticle size, protect the DNA from nucleases and other degrading enzymes during its transport in cell cytoplasm and nucleus and should not toxic to target cells. In this research, lipopeptide composed of palmitoyl (C-16) and short peptide sequence have been designed fully synthesized and tested to DNA condensation capability and toxicity. The DNA condensation study was performed using EtBr exclusion assay and cytotoxicity determination was carried out by colorimetric MTT assay. It was revealed that lipopeptide-based transfection agent of Pal-CKKHH and Pal-CKKHH-YGRKKRRQRRR-PKKKRKV condensed DNA molecules efficiently. The lipopeptide was less toxic compared to Lipofectamine and Poly-L-Lysine, that shown by 90% of CHO-K1 cells remained viable when they were treated with 4.36 µM Pal-CKKHHYGRKKRRQRRR-PKKKRKV. Meanwhile, there were only ~75% and 80% of CHO-K1 viable cells when it was treated with PLL and Lipofectamine®2000, respectively. Moreover, cell viability of HepG2 was ~ 75% after treated with 2.18 µM of Pal-CKKHH-YGRKKRRQRRR-PKKKRKV and decreased to ~65% when the lipopeptide concentration increased to 8.72 M. In summary, the synthesized lipopeptide condenses DNA molecules efficiently, less toxic than Lipofectamine®2000 and PLL and has possibility to be explored as a non-viral gene delivery vehicle.
Overproduction and Purification of Soluble Recombinant Human Granulocyte Colony Stimulating Factor in Escherichia coli Using Thioredoxin as Fusion Agustiyanti, Dian Fitria; Retnoningrum, Debbie Sofie; Rachmawati, Heni; Fuad, Asrul Muhamad
Annales Bogorienses Vol. 21 No. 1 (2017): Annales Bogorienses
Publisher : BRIN

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Recombinant human Granulocyte Colony Stimulating Factor (G-CSF) has been produced in a soluble form in Escherichia coli BL21 (DE3) as a fusion protein. The open reading frame of G-CSF was synthetically constructed in previous work and was codon optimized for best expression in E. coli. In this research, the gene was fused to thioredoxin (Trx) at the N-terminal in pET32 vector. The purpose of this research was to optimize the overproduction and purification processes to obtain high yield recombinant protein in soluble form, and to characterize the Trx-G-CSF fusion protein. Overproduction was performed using IPTG induction method for 3 and 6 hours. The protein was purified by Ni-NTA affinity chromatography and separated using gradient concentration of imidazole. The purified protein was then characterized by SDS-PAGE and Western Blot analysis. Further, enterokinase was used to separate G-CSF from the fusion protein. The purified form of G-CSF was subsequently characterized using Western Blot and mass spectrometry using MALDI-TOF. The results showed that the fusion protein was successfully produced in soluble part as much as 48.25% were obtained after 3 hours of induction. The yield of fusion protein was 67.37% from total protein (229.65 mg protein/L culture). The Western Blot analysis showed the G-CSF band at around 18.6 kDa. Mass spectrometry with MALDI-TOF/ TOF revealed that 25.86% of amino acid residue was recognized as part of human G-CSF sequence.
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 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 Pambudi, Sabar Pambudi, Sabar 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