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Corrigendum: Evaluating SARS-CoV-2 Spike Protein Transfection in HEK-293T Cells for VLP Applications Rohmah, Ilmiana Nurur; Marlita, Marlita; Kusuma, Kavana Hafil; Christina, Yuyun Ika; Dwijayanti, Dinia Rizqi; Mustikaningtyas, Dewi; Widodo, Nashi; Djati, M. Sasmito
The Journal of Experimental Life Science Vol. 15 No. 2 (2025)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jels.2025.015.02.01

Abstract

The spike protein of SARS-CoV-2 is crucial for initiating infections by binding to host cells and mediating membrane fusion. In this study, HEK-293T cells were transfected with plasmids encoding three structural proteins of SARS-CoV-2, i.e., Spike (S), Membrane (M), and Envelope (E). This transfection enabled the formation of SARS-CoV-2 Virus-Like Particles (VLPs), which allows for safer studies of the virus and its proteins. To serve as a marker for expression, an enhanced green fluorescent protein (EGFP) was fused to the spike protein, resulting in a Spike-EGFP (S-EGFP) fusion protein. The characteristics of SARS-CoV-2 spike protein expression in the transfected HEK-293T cells were then investigated using polymerase chain reaction (PCR) and flow cytometry. The PCR analysis revealed non-specific DNA band smearing, which did not provide conclusive confirmation of spike protein expression. However, flow cytometry analysis demonstrated that approximately 30% of the transfected cells exhibited green fluorescence, indicating the expression of the Spike-EGFP fusion protein. These findings, obtained through flow cytometry, confirmed the successful spike protein expression in transfected HEK-293T cells.  
Determination of Polyphenol Compounds in Clinacanthus nutans and Their Potential as Antioxidants Widodo, Nashi; Salma Wahid Marseti; Yuslinda Annisa; Fairuz Sarah Kamila; Rosyadah, Nuraini; Rifa’i, Muhaimin
Biotropika: Journal of Tropical Biology Vol. 13 No. 1 (2025)
Publisher : Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.biotropika.2025.013.01.06

Abstract

Free radicals can cause oxidative damage if they are produced in excess amounts in the body. To prevent this, the work of endogenous antioxidants needs to be supported by safe and effective exogenous antioxidants. Clinacanthus nutans is one of the sources of natural polyphenol compounds that have the potential to be used as free radical-scavenging agents. The study began with the extraction of C. nutans simplicia with microwave assisted extraction (MAE) using a 96% ethanol solvent. The amounts of phenols and flavonoids in the extract was then measured using the Folin-Ciocalteu and aluminium chloride techniques, and their antioxidant activity was assessed using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) scavenging assays. Based on the results of the study, it is known that C. nutans ethanolic extract contains higher flavonoids (238.29 ± 10.87 mgQE.g-1) than the total phenols (148.76 ± 1.26 mgGAE.g-1). This study also showed the potential of C. nutans extract as a free-radical scavenger with an IC50 from the NO-scavenging and DPPH assays of 28.37 ± 3.38 ppm and 117.84 ± 3.93 ppm, respectively. Despite the presence of high polyphenol content in the extract, its antioxidant activity was found to be relatively weak. Consequently, further study is necessary to ascertain the specific class of compounds responsible for its antioxidant activity and to assess its effectiveness.
Daruju (Acanthus ilicifolius L.) May Exhibit Anti-Breast Cancer Activity Through Inhibition of Proliferation Regulators: A Computational Study: Daruju (Acanthus ilicifolius L.) as Anti-Breast Cancer Agent Rosyadah, Nuraini; Kamila, Fairuz Sarah; Hermanto, Feri Eko; Widyananda, Muhammad Hermawan; Grahadi, Rahmat; Dwijayanti, Dinia Rizqi; Widodo, Nashi; Ulfa, Siti Mariyah
Journal of Tropical Life Science Vol. 15 No. 1 (2025)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/

Abstract

Breast cancer's increasing prevalence globally underscores the urgent need for effective and gentle therapies, positioning the exploration of herbal remedies as a critical pursuit. Daruju (Acanthus ilicifolius L.) emerges as a compelling candidate due to its inherent bioactive components. This research pioneers the application of advanced computational techniques to unveil the latent anti-breast cancer potential within A. ilicifolius. Our in-silico investigation commenced by cataloging A. ilicifolius compounds using the KNApSAcK database and existing literature. These compounds underwent rigorous screening for drug-like characteristics via SWISS-ADME and potential biological activity using PASS ONLINE. Protein targets relevant to breast cancer were predicted through SWISS Target and the STRING database, integrated with Cytoscape for network visualization. Molecular docking, performed with PyRx 0.8, assessed the binding strength between the identified compounds and target proteins, with the most promising interactions selected for further scrutiny. The stability of these crucial interactions was then evaluated through molecular dynamics simulations using YASARA. This comprehensive computational strategy aims to pinpoint potential anti-breast cancer agents derived from A. ilicifolius. Initial analysis of 17 compounds from A. ilicifolius, based on chromatography, databases, and prior studies, narrowed down to five that adhered to Lipinski’s Rule of Five for drug-likeness: 4-O-beta-D-glucosyl-4-coumaric acid, (-)-lyoniresinol, α-amyrin, adenosine, and p-coumaric acid. These compounds were predicted to directly interact with key breast cancer-related proteins across pathways like estrogen signaling, JAK/STAT, and PI3K/AKT. Notably, molecular docking revealed strong binding affinities for α-amyrin with CDK4, ER, and EGFR (-7.5 kcal/mol, -9.5 kcal/mol, and -8.7 kcal/mol, respectively), comparable to known inhibitors. Molecular dynamics simulations further corroborated the stability of these complexes, analyzing RMSD and binding affinity parameters. Consequently, α-amyrin stands out as a promising anti-breast cancer agent within A. ilicifolius, exhibiting potential to inhibit proteins crucial for breast cancer cell proliferation and survival, including CDK4, ER, and EGFR.
Design, Construction and Expression of Spike Highly Conserved Region (HCR) SARS-CoV-2 and Cholera Toxin Subunit B Fusion Protein in Lactococcus lactis NZ3900: Construction of recombinant plasmid in Lactococcus lactis Kesuma, Suryanata; Adianingsih, Oktavia Rahayu; Winarsih, Sri; Widodo, Nashi; Yurina, Valentina
Journal of Tropical Life Science Vol. 15 No. 2
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/2xf1qp96

Abstract

Prevention of SARS-CoV-2 transmission has primarily been achieved through vaccination, which is generally administered via injection and may cause discomfort. No commercially available SARS-CoV-2 vaccines can be administered via the mucosal route. However, recent advancements have demonstrated that vaccination with Lactococcus lactis enables vaccine delivery through the mucosa. A promising target for SARS-CoV-2 vaccine materials is the highly conserved region (HCR) of the SARS-CoV-2 spike (SARS-CoV-2 HCR Spike). Vaccine efficacy is enhanced by adding Cholera Toxin Subunit B (CTB) as an adjuvant. HCR and CTB proteins were recombinantly fused using a synthetic gene with optimized codons. This study aimed to construct a fusion protein of the SARS-CoV-2 spike protein and CTB in L. lactis strain NZ3900. The construction and expression of fusion proteins were analyzed using sequencing and protein electrophoresis. Codon optimization resulted in a Codon Adaptation Index value of 0.93 and a GC content of 27.06%. The cloning results revealed the formation of L. lactis colonies expressing the Fusion protein of the SARS-CoV-2 HCR Spike and CTB, which formed yellow colonies on the selection Elicker medium. PCR and sequencing confirmed the presence of the hcr-ctb gene, with a length of 981 bp and 100% sequence similarity. The Fusion protein of the SARS-CoV-2 HCR Spike and CTB was successfully expressed with a molecular weight of >35 kDa. In conclusion, we successfully constructed a Fusion protein of the SARS-CoV-2 HCR Spike and CTB in L. lactis NZ3900 as a potential vaccine candidate for oral administration to prevent SARS-CoV-2 infection.
Hepatoprotection and Immunomodulation of Natural Killer and CD8 T Cells by Meniran-Turmeric Extract Combination in Mice Injected with 7,12-Dimethylbenz[a] Anthracene Puspitarini, Sapti; Widodo, Nashi; Widyarti, Sri; Azzamuddin, Haidar; Ulinnuha, Maria; Puspitoputri, Alya H.; Khansa, Shafanony; Rifa’i, Muhaimin
HAYATI Journal of Biosciences Vol. 33 No. 2 (2026): March 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.2.499-507

Abstract

Combination herbal or polyherbal offers advantages by synergizing multiple plants to address health issues more effectively. Meniran and turmeric are traditionally used medicinal plants with notable pharmacological activities due to their bioactive compounds. This study investigated the curative effect of a meniran-turmeric extract combination on immunocompetent cells, particularly NK and CD8 cells, and on liver histopathology in mice administered 7,12-dimethylbenz[a]anthracene (DMBA). Female BALB/c mice (Mus musculus) were injected subcutaneously with DMBA at 45 mg/kg body weight (BW) in the mammary gland area and maintained for eight weeks. They were then treated with a 1:1 meniran-turmeric extract combination for two weeks at doses of 100, 300, 900, and 1,800 mg/kg BW, followed by liver and immune cell analyses. The results showed a lower CD8+ T-cell population in extract-treated groups (17.53±1.13%-19.79±1.51%) compared with the DMBA group (20.92±4.74%). Conversely, NK+ cell populations increased after extract treatment (0.33±0.01%-0.56±0.13%) compared with DMBA alone (0.27±0.02%). Liver histopathology indicated that extract combination administration did not cause more severe damage than that observed in the DMBA group. An extract combination of meniran and turmeric could modulate the immune system, particularly NK and CD8 cells, in mice injected with DMBA. In addition, the study indicated that the combination of meniran and turmeric extract might play a role as an antihepatotoxic agent. This research could serve as a consideration for future studies developing a combination formulation of meniran and turmeric.
Oral Immunization with Recombinant Lactococcus lactis and Retinoic Acid Boost Immune Response in Mice Nugroho, Inggil Cahyo; Amalia, Maulida Nisa; Juniardi, Muhamad Rizki; Marhendra, Eunike Berliana; Yanma, Nadifa Almira; Adianingsih, Oktavia Rahayu; Winarsih, Sri; Widodo, Nashi; Yurina, Valentina
Science and Technology Indonesia Vol. 11 No. 1 (2026): January
Publisher : Research Center of Inorganic Materials and Coordination Complexes, FMIPA Universitas Sriwijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26554/sti.2026.11.1.55-64

Abstract

The COVID-19 vaccine is one of the most important approaches for preventing SARS-CoV-2 virus transmission and spread. Oral vaccines are a promising option for preventing SARS-CoV-2 infection because it can activate both the mucosal and cellular immune systems. Previous research has shown that administering oral and intranasal vaccines can induce an immune response in mice. The effectiveness of the SARS-CoV-2 oral vaccine was evaluated in this study by combining spike protein with a carrier of food-grade recombinant Lactococcus lactis bacteria and a retinoic acid adjuvant. Mice were divided into three groups: a negative control (no treatment), a positive control (L. lactis recombinant), and a third group (L. lactis recombinant plus 300 μg retinoic acid). The vaccines were given three times, with a three-week interval between each. The serum levels of IgG, IgA, and IgE were determined using the ELISA method at the end of the study. CD4 and CD8 cellswere detected using immunofluorescence. While not statistically significant, the results showed that the retinoic acid groups had the highest anti-spike antibody levels of the three groups. In comparison to the control group, CD4 and CD8 cells increased in the spleens of mice given retinoic acid. There was no difference in temperature or IgE levels between vaccinated and non-vaccinated mice, indicating that the vaccine caused no allergic reaction. This study’s findings suggest that retinoic acid adjuvant can stimulate the cellular and humoral immune system.
Development of a Stable CHO Cell Line for High-Level Expression and Purification of Recombinant Darbepoetin Alfa Widekdo, Dwi purno; Widodo, Nashi; Rifa'i, Muhaimin; Jatmiko, Yoga Dwi
Journal of Tropical Life Science Vol. 15 No. 3
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.15.03.12

Abstract

Recombinant Darbepoetin alfa (DARB), an erythropoiesis-stimulating agent, is widely used in treating anemia linked to chronic kidney disease and chemotherapy. This study presents a systematic approach for developing a high-yield, stable Chinese hamster ovary (CHO) cell line capable of producing recombinant DARB. A codon-optimized gene construct under dual CMV/EF1α promoters and puromycin resistance selection was transfected into CHO cells. To isolate high-producing monoclonal populations, transfected cells were subjected to limiting dilution cloning in 96-well plates, allowing single-cell-derived colonies to expand and be individually assessed. Clone DARV IV Pool 2 Clone 2E6 demonstrated the highest and most consistent DARB expression, validated through Western blot using anti-human EPO antibodies. The culture supernatant underwent two-step clarification via centrifugation and tangential flow filtration (TFF), followed by purification using anion-exchange chromatography on a HiTrap Q HP column. Gradient elution enabled effective separation, with SDS-PAGE and Western blot confirming high purity and molecular integrity of the recombinant protein. Variability in clone expression highlighted the influence of genomic integration sites and potential epigenetic silencing, emphasizing the importance of screening and stability validation. This study demonstrates that rational vector design, antibiotic-based clone selection, and robust purification strategies can produce CHO-derived DARB suitable for large-scale production. The integrated workflow supports scalability, product consistency, and regulatory readiness for biosimilar therapeutic manufacturing
The Role of Moringa oleifera- Ifalmin® Formulation in Regulation of B220+IgM+ and B220+IgG+ in Diabetic Mice Adharini, Wahyu Isnia; Nilamsari, Ruri Vivian; Lestari, Noviana Dwi; Widodo, Nashi; Rifa'i, Muhaimin
The Journal of Experimental Life Science Vol. 10 No. 1 (2020)
Publisher : Graduate School, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (924.127 KB) | DOI: 10.21776/ub.jels.2019.010.01.08

Abstract

It has been known that the immunoglobulin levels were altered in diabetes mellitus (DM) conditions. This study aimed to evaluate the levels of immunoglobulins in DM mice after the administration of Moringa oleifera-Ifalmin® formulation (MI). Streptozotocin, at a dose of 145 mg.kg-1, was injected intraperitoneally to experimental mice to obtain diabetic mice. The groups were divided into normal mice, diabetic mice without treatment, diabetic mice with metformin treatment (307.5 mg.kg-1 BW), and diabetic mice with MI treatment at dose 1 (M:I= 800 mg.kg-1 BW: 800 mg.kg-1 BW), dose 2 (M:I= 615 mg.kg-1 BW: 615 mg.kg-1 BW), and dose 3 (M:I= 800 mg.kg-1 BW: 615 mg.kg-1 BW). Mice were orally treated by MI for 14 days. Subsequently, the levels of immunoglobulin IgM and IgG were evaluated using flow cytometry analysis. IgM and IgG levels were significantly lower in the DM group than the normal group. These results indicated that DM altered immunoglobulin levels. MI treatment for 14 days significantly increased the number of IgM and IgG at the level equivalent to the normal group and significantly different as compared to the DM group. Based on the results, MI can be used as an immunomodulatory agent in humoral immunity through the precise regulation of IgM and IgG.
Co-Authors Achmad Ridok Adharini, Wahyu Isnia Adi Hermansyah, Adi Agatha Mia Puspitasari Agatha Mia Puspitasari Amalia, Maulida Nisa Annisa, Yuslinda Aris Soewondo Azerlyn, Defiona Rensia Naomi Azzamuddin, Haidar Custer C. Deocaris Deocaris, Custer C. Dewi Mustikaningtyas Didik Priyandoko Didik Priyandoko Dinia Rizqi Dwijayanti Dodi Iskandar, Dodi Dwi Adi Nugroho Fairuz Sarah Kamila Fitria Eka Aprilia Grahadi, Rahmat Hafil Kusuma, Kavana Hermanto, Feri Eko Ichsan Ichsan Ika Christina, Yuyun Ikhtiarini, Nur Inggita Kusumastuty Johan Aloysius Juniardi, Muhamad Rizki Kamila, Fairuz Sarah Kaul, Sunil C. Kesuma, Suryanata Khansa, Shafanony Kholilah, Tsaniyah Nur Kusuma, Kavana Hafil Lestari, Noviana Dwi Listijani Suhargo, Listijani Marhendra, Eunike Berliana Mariyah Ulfa, Siti Marlita Marlita, Marlita Masruri Masruri Mifetika Lukitasari Moch Sasmito Djati Mohammad Saifur Rohman Nafisah, Wirdatun Nia Kurniawan Nilamsari, Ruri Vivian Nugroho, Inggil Cahyo Nur Ida Panca Nugrahini Oktavia Rahayu Adianingsih Puspitoputri, Alya H. raudatul jannah raudatul jannah Renu Wadhwa Rifa'i, Muhaimin Rifa’i, Muhaimin Rizqi Dwijayanti, Dinia Rohmah, Ilmiana Nurur Rollando, Rollando Rosyadah, Nuraini ROUDLOTUL JANNAH Roudlotul Jannah Salma Wahid Marseti Sapti Puspitarini Siti Fatiyatur Rahmah Siti Fatiyatur Rahmah Siti Mariyah Ulfa Siti Nurkasanah Sri Widyarti Sri Winarsih Sunil C. Kaul Sutiman B. Sumitro Ulinnuha, Maria VALENTINA YURINA Wadhwa, Renu Warsito Warsito Wayan Firdaus Mahmudy Widekdo, Dwi purno Widyananda, Muhammad Hermawan Yanma, Nadifa Almira YOGA DWI JATMIKO Yuslinda Annisa Yuyun Ika Christina