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All Journal ANNALES BOGORIENSES JFIOnline
Endah Puji Septisetyani, Endah Puji
Research Center for Biotechnology Indonesian Institute of Sciences Jl. Raya Bogor KM 46, Cibinong, Bogor, Indonesia 16911
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Health Professions Immunology & microbiology Medicine & Pharmacology

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Sequential Adaptation in Mammalian CHO-K1 Cells Producing Human Erythropoietin Wisnuwardhani, Popi Hadi; Septisetyani, Endah Puji; Santoso, Adi
ANNALES BOGORIENSES Vol 21, No 1 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (289.139 KB) | DOI: 10.14203/ab.v21i1.282

Abstract

The production of recombinant proteins for clinical applications using mammalian cell technology has become a prevalent system because of its capacity in assembling functional proteins.  One of the main problems with CHO-K1 cells is that this cell has to grow in the presence of serum. However, the presence of serum will complicate the downstream step for protein production. Thus, protein produced in media without serum, theoretically, would be easier to purify.  Technically, this type of cell can be produced by growing the CHO-K1 cells in serum-free media by using adaptation method in suspension condition. This research showed that through sequential adaptation using conditioned media, the CHO-K1 cell line that produces the human erythropoietin gene (hEPO) was able to grow in suspension culture using serum-free media.  Based on Western blot analysis, it showed that the protein (hEPO) was able to be expressed in suspension culture with molecular mass of about 47 kDa.
Optimizaton of Cationic Lipid Mediated Transfection of pEGFP-c1 and pJ-EPO Plasmids in Chinese Hamster Ovary (CHO) Cells Attached Culture for Transient and Stable Recombinant Human Erythropoietin (rhEPO) Expression Septisetyani, Endah Puji; Kusumawati, Arizah; Santoso, Adi
ANNALES BOGORIENSES Vol 19, No 1 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v19i1.84

Abstract

Cationic lipid is one of transfection agents which show high efficiency and low cytotoxicity. The transfection efficiencies may depend upon the type or amount of cationic lipids, the cell line or DNA plasmid being used for transfection. The purpose of this study was to find optimal condition for transfection of CHO-K1 and CHO-S cells with pJ-EPO plasmid (contain human epo gene) compared with pEGFP-c1 plasmid (contain gfp gene) by cationic lipid Lipofectamin 2000TM to generate stable transfectant expressing recombinant human erythropoietin (rhEPO). Optimization was carried out regarding the amount of lipofectamin, DNA concentration, and concentration of antibiotic Geneticin (G418) for selection of stable transfectants. Using standard amount of lipofectamin (10µl/well) in 6-well plate, highest expression level of green fluorescent protein (GFP) was shown after transfection of CHO-K1 cells with 4µg/well pEGFP-c1 while highest expression level of rhEPO was observed after transfection of CHO-K1 cells with 6, 8, and 10µg/well pJ-EPO plasmids. The data also indicated that optimal transfection conditions of CHO-K1 and CHO-S cells with pJ-EPO were shown with the use of 4µg/well DNA and 15µl lipofectamin, respectively. Concentration of G418 affected the expression where strongest rhEPO expression was shown at 1,000 ng/µl G418 concentration. 
8-HIDROKSIISOKAPNOLAKTON-2',3'-DIOL, KUMARIN BIOAKTIF DARI Micromelum minutum Susidarti, Ratna Asmah; Rahmani, Mawardi; Sukari, Mohd. Aspollah; Ali, Abdul Manaf; Mustofa, .; Yasmina, Alfi; Handayani, Sri; Mintarsih, Betty; Ikawati, Muthi’; Septisetyani, Endah Puji
JFIOnline | Print ISSN 1412-1107 | e-ISSN 2355-696X Vol 4, No 3 (2009)
Publisher : Indonesian Research Gateway

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Abstract

Separation of leaves chloroform extract of Micromelum minutum (Rutaceae) yielded a new coumarin, 8-hydroxyisocapnolactone-2¢,3¢-diol. The structure of this compound was characterized by UV, IR, MS and NMR spectroscopic methods, including 1H, 13C, HSQC, COSY, HMBC dan NOESY experiments. This compound is significantly toksisk towards several cancer cell lines (CEM-SS, HL60, HeLa, HepG2, MCF7, T47D and NS1), active against chloroquin sensitive (D10) and resistance (FCR3) Plasmodium falciparum and showed strong antibacterial activity against Bacillus Subtilis mutan, Bacillus. Subtilis wild type, Pseudomonas Aeruginosa dan Staphylococcus aureus resisten meticilin).  ABSTRAK Pemisahan ekstrak kloroform daun Micromelum minutum (Rutaceae) menghasilkan suatu kumarin baru, 8-hidroksiisokapnolakton-2΄,3΄-diol yang strukturnya diidentifikasi secara spektroskopi UV, IR, MS dan NMR termasuk 1H, 13C, HSQC, COSY, HMBC dan NOESY. Senyawa tersebut secara signifikan toksik terhadap beberapa sel kanker (CEM-SS, HL60, HeLa, HepG2, MCF7, T47D dan NS1), aktif terhadap Plasmodium falciparum yang sensitif (D10) maupun resisten (FCR3) kloroquin dan mempunyai aktivitas antibakteri yang kuat terhadap Bacillus Subtilis mutan, Bacillus. Subtilis wild type, Pseudomonas Aeruginosa dan Staphylococcus aureus resisten meticilin).
8-HIDROKSIISOKAPNOLAKTON-2',3'-DIOL, KUMARIN BIOAKTIF DARI Micromelum minutum Susidarti, Ratna Asmah; Rahmani, Mawardi; Sukari, Mohd. Aspollah; Ali, Abdul Manaf; Mustofa, .; Yasmina, Alfi; Handayani, Sri; Mintarsih, Betty; Ikawati, Muthiâ??; Septisetyani, Endah Puji
Jurnal Farmasi Indonesia Vol 4, No 3 (2009)
Publisher : Jurnal Farmasi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35617/jfi.v4i3.18

Abstract

Separation of leaves chloroform extract of Micromelum minutum (Rutaceae) yielded a new coumarin, 8-hydroxyisocapnolactone-2¢,3¢-diol. The structure of this compound was characterized by UV, IR, MS and NMR spectroscopic methods, including 1H, 13C, HSQC, COSY, HMBC dan NOESY experiments. This compound is significantly toksisk towards several cancer cell lines (CEM-SS, HL60, HeLa, HepG2, MCF7, T47D and NS1), active against chloroquin sensitive (D10) and resistance (FCR3) Plasmodium falciparum and showed strong antibacterial activity against Bacillus Subtilis mutan, Bacillus. Subtilis wild type, Pseudomonas Aeruginosa dan Staphylococcus aureus resisten meticilin).  ABSTRAK Pemisahan ekstrak kloroform daun Micromelum minutum (Rutaceae) menghasilkan suatu kumarin baru, 8-hidroksiisokapnolakton-2Î?,3Î?-diol yang strukturnya diidentifikasi secara spektroskopi UV, IR, MS dan NMR termasuk 1H, 13C, HSQC, COSY, HMBC dan NOESY. Senyawa tersebut secara signifikan toksik terhadap beberapa sel kanker (CEM-SS, HL60, HeLa, HepG2, MCF7, T47D dan NS1), aktif terhadap Plasmodium falciparum yang sensitif (D10) maupun resisten (FCR3) kloroquin dan mempunyai aktivitas antibakteri yang kuat terhadap Bacillus Subtilis mutan, Bacillus. Subtilis wild type, Pseudomonas Aeruginosa dan Staphylococcus aureus resisten meticilin).
Co-Authors Abdul Manaf Ali, Abdul Manaf Alfi Yasmina Kusumawati, Arizah Mintarsih, Betty Mintarsih, Betty Mustofa Mustofa Muthi Ikawati Popi Hadi Wisnuwardhani, Popi Hadi Rahmani, Mawardi Rahmani, Mawardi Ratna Asmah Susidarti Sri Handayani Sukari, Mohd. Aspollah Sukari, Mohd. Aspollah