Popi Hadi Wisnuwardhani, Popi Hadi
Research Center for Biotechnology, Indonesian Institute of Sciences Jalan Raya Bogor km 46 Cibinong, West Java, Indonesia, 16911

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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.
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

Abstract

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.
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

Abstract

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

Abstract

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

Abstract

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.
Mini Review: GLP-1 Modification, Development, and Improvement Damai, Fedric Intan; Purwanto, Gracia Christine Lembong; Wardiana, Andri; Wisnuwardhani, Popi Hadi; Agustiyanti, Dian Fitria; Fathurahman, Alfi Taufiq; Ningrum, Ratih Asmana
Annales Bogorienses Vol. 26 No. 1 (2022): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2022.v26.n1.1-12

Abstract

Diabetes Mellitus Type 2 (DM-2) is the condition where the body comes to be insensitive or even resistant towards insulin, thus resulting in deficient insulin secretion from beta cells in the pancreas. Compared with the available treatments, Glucagon-like peptide 1 (GLP-1) is considered a natural treatment to cure DM-2 due to its characteristic as an incretin hormone, where one of its functions is to improve insulin secretion and enhance beta-cell glucose sensitivity. However, GLP-1 has a limitation, which is a rapid half-life due to active degradation activities in the body. Therefore, many studies have been conducted to develop and improve the pharmacological activity of GLP-1 through structural modification and yield improvement, which are thoroughly reviewed in this paper. Structural modification of GLP-1 covers amino acid substitutions by referring to the GLP-1 analog, Exendin4, to prevent the dipeptidyl peptidase-4 (DPP-4) degradation activity and protein fusion with an additional chain to extend the half-life during administration. The yield improvement at the overexpression of GLP-1 tandem repeats sequences can increase the transcribed genes’ yield. The studies show that specific amino acid substitutions and IgG heavy chain constant regions or Fc-based fusion genes successfully enhance the pharmacological activities of GLP-1. At the same time, Pichia pastoris expression system managed to yield 219.2 mg/l target protein, where the purified target protein is capable of producing 10× yield of a single GLP analog. Further research could include the utilization of these findings in vitro as a GLP-1 analog-based therapeutics to treat DM-2.