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The Emergence of Biosimilars in Indonesia: Guidelines, Challenges and Prospects Wardiana, Andri; Ningrum, Ratih Asmana
ANNALES BOGORIENSES Vol 20, No 2 (2016): 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.v20i2.272

According to the Food and Drug Administration (FDA), biosimilar is defined as a product which is highly similar to the reference product without clinically meaningful differences in safety, purity and potency. Indonesia is a developing country which has more than 250 million people. The domestic need of Biosmilar in Indonesia is about 10-20%.Â Even though the need is very high, Indonesia still has not been able to produce Biosimilar independently. To stimulate the domestic production on biosimilar, National Agency for Drug and Food Control (NADFC) Republic of Indonesia has assigned Regulation of Biosimilar as Peraturan Kepala Badan Pengawas Obat Dan Makanan Republik Indonesia Nomor 17 Tahun 2015 Tentang Pedoman Penilaian Produk Biosimilar. The guidance covers the quality requirement and evaluation of Biosimilar products. The Ministry of Health of Indonesia has a strategic plan in biopharmaceutical covering biosimilar which is going to develop in 2015-2025. The strategy is expected to initiate biosimilar production in Indonesia. This review focuses on the guidelines, challenges and prospects biosimilars in Indonesia comparing to other international regulatory bodies.Keywords: Biosimilar, Indonesia, Guidelines, Challenges and Prospects

Roferon-A: A Biologic Product of Human Interferon Alpha 2a Wardiana, Andri; Ningrum, Ratih Asmana
ANNALES BOGORIENSES Vol 19, No 2 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Human interferon alpha 2a (hIFNÎ±2a) is a cytokine regulating immune system that has been used in hepatitis and cancer treatments. It has wide biological potency covering antiviral, antiproliferative and immunomodulative activities. This mini review discusses Roferon-A as a prominent commercial product of recombinant hIFNÎ±2a which is produced in bacterial system, Escherichia coli, as therapeutic protein for several diseases, such as chronic viral Hepatitis B, Hepatitis C, melanoma, hairy cell leukemia and renal cell carcinoma. The discussion focuses on the development process with regard to its manufacturing, preclinical and clinical studies, as well as therapeutic efficacy. In addition, we also discuss biosimilar development of hIFNÎ±2a and its potential future developments in the context of enhancing pharmacokinetic profiles

Expression of No Affinity Tagged Recombinant Human Interferon Alpha-2a in Methilotropic Yeast Pichia pastoris Herawati, Neng; Wardiana, Andri; Ningrum, Ratih Asmana
ANNALES BOGORIENSES Vol 19, No 2 (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.v19i2.254

Recombinant human interferon alpha-2a (rhIFNÎ±-2a) has been widely used for clinical therapy as antiviral, anticancer as well as immunomodulator. In this study, the open reading frame (ORF) encoding synthetic hIFNÎ±-2a was constructed to be in framed with N-terminal alpha factor secretion system in methylotropic yeast Pichia pastoris. This research aimed to construct, express and analyse the non-affinity tagged recombinant human interferon alpha-2a in the methilotropic yeast P. pastoris. We used pPICZÎ±B plasmid for cloning and expression vector. The confirmed recombinant plasmid containing the correct DNA sequence of hIFNÎ±-2a was linearized by SacI restriction enzyme, then transformed into P. pastoris genome using electroporation. We screened two multi-copy recombinants in YPDS plates containing Zeocinâ¢. Buffered complex medium containing 0.5 % methanol (BMMY) was used for protein expression for 48 hours in the culture condition. The recombinant protein was purified by blue sepharose affinity chromatography. Analyses of hIFNÎ±-2a protein by SDS-PAGE and Western blot confirmed that protein band in which was observed around 19.2 kDa, was recombinant hIFNÎ±-2a. The quantification of purified rhIFNÎ±-2a using colorimetric binichoninic assay (BCA) informed that the yield was 44 mg/L culture (OD600= 2-3).

Optimization of Expression Condition, Two Dimensional And Melting Point-Based Characterization of Recombinant Human Interferon Alpha-2a Fusion and Non Fusion Forms Ningrum, Ratih Asmana; Wardhani, Widdya Kusuma; Wahyuni, Ike; Mustopa, Apon Zaenal
ANNALES BOGORIENSES Vol 22, No 2 (2018): 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/ann.bogor.2018.v22.n2.57-64

Recombinant Human Interferon Alpha-2a (rhIFNα-2a) is a therapeutic protein that used in hepatitis and cancer treatments. In our previous research, we developed higher molecular weight of the protein through human serum albumin fusion. The fusion and non fusion form of rhIFNα-2a were produced in Pichia pastoriswith 86 kDa and 19 kDa in size respectively. In previous research, protein yield was not reproducible due to unoptimized expression conditions. This reseach was aimed to optimize expression condition process and to characterize the fusion and non fusion forms of rhIFNα-2a. The parameters to observe in overproduction include nutrient (media and methanol concentration) and non nutrient (temperature andincubation period). Affinity and size exclusion cromatographicwere compared in protein purification. BCA assay was used to determine quantity of protein. Protein characterization was conducted using two-dimensional SDS PAGE and denaturation analyses. The optimal condition of expression was achieved using complex media with 1% of methanol for 3 day incubation period at 25°C. The protein yield was reproducible and higher comparing to previous research. Affinity chromatography resulted in higher purity of the proteins comparing to size exclusions. Characterization using two dimensional gel analysis revealed that isoelectric point of rhIFNα-2a is 6.5 for fusion form and 6.0 for non fusion form. The melting points of fusion protein were 56°C and 62°C whilst that of non fusion was 56°C.

Naringin Effect on SARS-CoV-2 Pseudovirus Entry and Spike Mediated Syncytia Formation in hACE2-overexpressing Cells Septisetyani, Endah Puji; Prasetyaningrum, Pekik Wiji; Paramitasari, Komang Alit; Suyoko, Ahmad; Himawan, Alayna Lillahida Indri; Azzahra, Salsabila; Wisnuwardhani, Popi Hadi; Anam, Khairul; Ramadani, Ratna Dwi; Santoso, Adi; Ningrum, Ratih Asmana; Herawati, Neng; Rubiyana, Yana
HAYATI Journal of Biosciences Vol. 31 No. 2 (2024): March 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.2.336-347

A molecular docking study demonstrates the interaction between naringin, a citrus flavonoid, with SARS-CoV-2 spike RBD. Nevertheless, in vitro investigation of the inhibitory effect of naringin on SARS-CoV-2 entry and syncytia models has yet to be carried out. We synthesized VSV∆G-GFP/Spike* pseudovirus (PSV) as a SARS-CoV-2 model by pseudotyping VSV∆G-GFP/S* in BHK-21 cells overexpressing the SARS-CoV-2 spike glycoprotein. In the SARS-CoV-2 PSV entry assay, we utilized CHO-K1 cells transfected with hACE2 plasmid, which were then treated with naringin and SARS-CoV-2 PSV/naringin. After 16-18 h incubation, PSV internalization represented by the GFP signal was observed under a fluorescence microscope. Immunofluorescence staining was also performed to probe the SARS-CoV-2 spike and confirm the PSV entry. We performed a syncytia assay using 293T cells co-transfected with SARS-CoV-2 spike/hACE2. Six hours after transfection, the cells were treated with naringin and incubated for another 16-18 hours. Then, we observed syncytia using a phase contrast microscope. Based on fluorescence foci quantification, the results indicated that naringin might inhibit SARS-CoV-2 PSV entry at a concentration of 100 µM (P<0.05). However, naringin did not prevent syncytia formation compared to solvent control. These PSV entry and syncytia assay results suggested that naringin potentially inhibited SARS-CoV-2 viral infection but not cell-to-cell viral transmission.

Medium Optimization for Recombinant Human Papillomavirus Type 52 L1 Protein Production in Pichia pastoris GS115 Platform on Bioreactor Scale Mustopa, Apon Zaenal; Nur Amani, Febriyanti; Irawan, Herman; Novianti, Ela; Swasthikawati, Sri; Ekawati, Nurlaili; Nurfatwa, Maritsa; Joko Wahyono, Daniel; Juanssilfero, Ario Betha; Mamangkey, Jendri; Purnomo, Yudi; Hertati, Ai; Wijaya, Hans; Dewi, Kartika Sari; Ningrum, Ratih Asmana
HAYATI Journal of Biosciences Vol. 32 No. 5 (2025): September 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.5.1283-1294

Human papillomavirus (HPV) stands as the primary etiological agent in the development of invasive cervical cancer worldwide. The L1 protein is a pivotal constituent of prophylactic HPV vaccines. Notably, HPV type 52 is one of the most prevalent genotypes found in squamous cell carcinoma cases in Indonesia. This research endeavor aims to enhance the productivity of recombinant HPV-52 L1 protein by optimizing the culture conditions of P. pastoris GS115 cells. In this study, we conducted trials employing 17 different media variants to optimize the expression of recombinant HPV-52 L1 protein. The results from small-scale experiments revealed three media, namely SYN6.10, BMMY, and SYN6.1, which exhibited promising yields of recombinant HPV-52 L1 protein as assessed through ELISA or immunoassay analysis. We succeeded in refining the SYN6.10 derivative, denoted as SYN6.10b, specifically designed for use in 1-L and 5-L bioreactors. This achievement was realized by adjusting Trace Element Solution (TES) and Vitamin Solution (VS) concentrations and implementing a methanol fed-batch phase with the addition of 0.3% methanol after 24 and 48 hours of fermentation in the P. pastoris medium. Further visualizations through SDS-PAGE and western blot analysis confirmed the protein after 72 hours of fermentation in a 1-L bioreactor using the SYN6.10b medium. In conclusion, the SYN6.10b medium required a 72 hours fermentation period to successfully express recombinant HPV-52 L1 protein in the P. pastoris platform.

Diagnostic Value of Saliva RT-PCR Test within Suspected SARS-CoV-2 Cases in Indonesia Putra, Andika Chandra; Zaini, Jamal; Ridwanuloh, Asep Muhammad; Nugroho, Herjuno Ari; Setyawan, Ryan Haryo; Idris, Idris; Setiawan, Ruby; Sushadi, Pangda Sopha; Wulandari, Ari Sulistyo; Zannati, Anky; Indriawati, Indriawati; Erdayani, Eva; Wahyuni, Wahyuni; Agustiyanti, Dian Fitria; Wisnuwardhani, Popi Hadi; Saniyyah, Zahrah; Azika, Wira Norman; Haryanto, Budi; Utomo, Ahmad Rusdan Handoyo; Ningrum, Ratih Asmana
Health and Medical Journal Vol 6, No 2 (2024): HEME May 2024
Publisher : Universitas Baiturrahmah

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33854/heme.v6i2.1494

Introduction: The ongoing SARS-CoV-2 pandemic has profoundly emphasized the pressing need for accurate and reliable diagnostic procedures. Given the potential health risks associated with nasopharyngeal swabs, there has been growing interest in seeking alternative diagnostic mediums. In this context, our study delved into evaluating saliva as a potential diagnostic tool, simultaneously assessing its efficiency in relation to patient demographics and their exhibited clinical symptoms. Methods: Spanning from May to December 2020, we conducted a comprehensive cross-sectional analysis. We meticulously examined medical records to gather insights on patient characteristics, existing health conditions, onset of symptoms, clinical manifestations, and compared the results obtained from both salivary and nasopharyngeal RT-PCR tests for SARS-CoV-2. Results: Among the individuals suspected of SARS-CoV-2 infection, the mean age stood at 52.4 years, with males representing 60.3% of this group. Interestingly, a significant 76.9% reported underlying health conditions, predominantly hypertension and diabetes. The most commonly reported symptoms encompassed respiratory challenges, notably coughing and shortness of breath, succeeded by symptoms like nausea, fever, and a general sense of fatigue. The performance of saliva tests, in terms of accuracy, appeared to be significantly influenced by the timing of symptom emergence. Conclusion: The RT-PCR tests utilizing saliva samples demonstrated considerable promise, especially during the early stages of symptom manifestation, providing a reliable alternative to traditional nasopharyngeal swabs. The findings suggest a superior diagnostic sensitivity when utilizing saliva during the initial phases of a SARS-CoV-2 infection.

Antioxidant and Cytotoxic Activities of Lactic Acid Bacteria on Colorectal Cancer WiDr Cell Line Wisnuwardhani, Popi Hadi; Ningrum, Ratih Asmana; Mustopa, Apon Zaenal; Vanggi, Leggina Rezzy; Kusdianawati, Kusdianawati Kusdianawati
Indonesian Journal of Cancer Chemoprevention Vol 12, No 1 (2021)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev12iss1pp28-36

Colorectal cancer (CRC) is one of the leading causes of cancer and cancer-related deaths worldwide. Lactic acid bacteria (LAB) are bacteria that have potential activity as an inhibitor of the growth of colorectal cancer, and also has been widely used and was very useful for consumption. In our previous study, we isolated various LAB from Indonesian traditional fermented food. This study aims to determine the potential of LAB as an anticancer agent by determining the antioxidant activity and cytotoxicity assay of colon cancer in the WiDr cell line. This study used extracellular extract of various LAB. We use the Diphenylpicrylhydrazyl (DPPH) method to determine the antioxidant activity and 3-(4,5'dimethylihiazol-2-yl),2.5-di-phenyl-relrrzolium bromid (MTT) assay to study cytotoxicity activity. The viability cell staining also applied to detect unviable cells. The results informed that the highest antioxidant activity was shown by S.34 LAB with 81% activity. The S.34 also showed cytotoxicity activity with 73% of WiDr viable cell at a concentration of 200 μg/mL of LAB extract. Based on the results of the study, it can be concluded that the S.34 LAB from Bekasam may inhibit the proliferation of WiDr cell lines and It had the highest antioxidant activity comparing to other LAB samples.Keywords: Lactic Acid Bacteria, colorectal cancer, anticancer, antioxidant, WiDr cells.

Evaluation of Curcumin-derived Carbon-dots' Inhibitory Activity as SARS-CoV-2 Antiviral Candidate Using Chemical Crosslinking Taharuddin, Audrey Angelina Putri; Yamahoki, Nicholas; Stephanie, Rebecca; Agustiyanti, Dian Fitria; Wisnuwardhani, Popi Hadi; Angelina, Marissa; Rubiyana, Yana; Ningrum, Ratih Asmana; Wardiana, Andri; Desriani, Desriani; Hariyatun, Hariyatun; Iskandar, Ferry; Permatasari, Fitri Aulia; Giri-Rachman, Ernawati Arifin; Fibriani, Azzania
HAYATI Journal of Biosciences Vol. 33 No. 1 (2026): January 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.1.232-239

In our previous work, we demonstrated that curcumin-derived carbon dots (Cur-CDs) have potential as antivirals for COVID-19. However, the precise mechanism of action remains unclear. This study investigated the potential of Cur-CDs against SARS-CoV-2 by targeting the dimerization of the C-terminal domain of nucleocapsid protein (N-CTD) using chemical crosslinking. Recombinant SARS-CoV-2 N-CTD was expressed, purified, and subjected to chemical crosslinking. The dimerization inhibition ability of Cur-CDs was assessed with ligand concentrations ranging from 0 to 2,000 μg/mL. Successful inhibition —defined as a noticeable reduction in SARS-CoV-2 N-CTD dimer band intensity on SDS-PAGE—was observed when Cur-CDs were present at 8 to 16 times the protein concentration. We hypothesize that Cur-CDs bind to the dimerization residues, preventing non-covalent interactions between monomers and limiting dimer formation. Our findings suggest that Cur-CDs could be a promising antiviral strategy for SARS-CoV-2, especially targeting the dimerization of the nucleocapsid protein. Additionally, this study also highlights the use of chemical crosslinking as a valuable tool for interaction-based drug screening.

Optimization of Expression Condition, Two Dimensional And Melting Point-Based Characterization of Recombinant Human Interferon Alpha-2a Fusion and Non Fusion Forms Ningrum, Ratih Asmana; Wardhani, Widdya Kusuma; Wahyuni, Ike; Mustopa, Apon Zaenal
Annales Bogorienses Vol. 22 No. 2 (2018): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar

Recombinant Human Interferon Alpha-2a (rhIFNα-2a) is a therapeutic protein that used in hepatitis and cancer treatments. In our previous research, we developed higher molecular weight of the protein through human serum albumin fusion. The fusion and non fusion form of rhIFNα-2a were produced in Pichia pastoriswith 86 kDa and 19 kDa in size respectively. In previous research, protein yield was not reproducible due to unoptimized expression conditions. This reseach was aimed to optimize expression condition process and to characterize the fusion and non fusion forms of rhIFNα-2a. The parameters to observe in overproduction include nutrient (media and methanol concentration) and non nutrient (temperature andincubation period). Affinity and size exclusion cromatographicwere compared in protein purification. BCA assay was used to determine quantity of protein. Protein characterization was conducted using two-dimensional SDS PAGE and denaturation analyses. The optimal condition of expression was achieved using complex media with 1% of methanol for 3 day incubation period at 25°C. The protein yield was reproducible and higher comparing to previous research. Affinity chromatography resulted in higher purity of the proteins comparing to size exclusions. Characterization using two dimensional gel analysis revealed that isoelectric point of rhIFNα-2a is 6.5 for fusion form and 6.0 for non fusion form. The melting points of fusion protein were 56°C and 62°C whilst that of non fusion was 56°C.

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