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All Journal HAYATI Journal of Biosciences Makara Journal of Science Annales Bogorienses
Yana Rubiyana, Yana
Research Center for Biotechnology, Indonesian Institute of Sciences Jl. Raya Bogor KM 46, Cibinong, Bogor, 16911, Indonesia
Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences

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 1 
Comparison of Immobilized Metal Affinity Chromatography Ni-NTA and Co-TALON for the Purification of Recombinant Human Erythropoietin Rubiyana, Yana; Santoso, Adi; Batubara, Irmanida
Makara Journal of Science Vol. 19, No. 4
Publisher : UI Scholars Hub

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Abstract

The purification of recombinant proteins is an important stage in biopharmaceutical research. A commonly used technique is immobilized metal affinity chromatography (IMAC). One of the main advantages of this type of chromatography is that the column can easily be regenerated for subsequent purification work. The mechanism of IMAC is based on bonding between metal ions immobilized on a matrix with a specific amino acid. Because of the strong interactions of the electron donor group on the imidazole ring, histidine is often used in the IMAC purification system. Two types of commercial IMAC resin use a nitrilotriacetic acid (NTA) matrix: a nickel-based (Ni-NTA) and cobalt-based (Co-NTA), better known as TALON. This study was aim to investigate the effect of the metal ions Ni2+ and Co2+ to purify recombinant human erythropoietin (rhEPO) expressed in yeast system Pichia pastoris. The results indicated that both Ni-NTA and Co-TALON gave almost the same level of protein purity; however, Ni-NTA has a higher binding affinity than Co-TALON might be due to the higher stability complex of Ni+. The average amount of protein bound by Ni-NTA and Co-TALON was 183.5 and 38.7 µg/mL, respectively.
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.
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.
In-Silico Cloning and Analysis of Divalent Subunit OMP31-SODc Proteins As A Prophylaxis Vaccine Against Brucella melitensis Infection Wijaya, Sri Kartika; Kusumawati, Arizah; Wardiana, Andri; Rubiyana, Yana; Husnaa, Ulfatul; Santoso, Adi
Annales Bogorienses Vol. 19 No. 1 (2015): Annales Bogorienses
Publisher : BRIN

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Abstract

The urgency to develop a new protein subunit based vaccine candidate against Brucella was provoked by its frequent infection to human and livestock. Since Brucella melitensis is found as the most frequently isolated Brucella species from human, thus the outer membrane of B. melitensis becomes a prominent subcellular localization to search for promising antigen to be developed as vaccine candidate due to its interaction with host cell. Among outer membrane proteins suggested by Vaxign program, OMP31 was found as the most promising candidate. Moreover, analysis on other subcellular localization led our interest to SODc protein, which was expected to support OMP31 in triggering immune response. The OMP31-SODc divalent vaccine candidate was analysed in silico to predict its stable three-dimensional structure, cloning process and expectation on the ease during expression, purification and vaccine delivery to elicit the expected immune response.
Heterologous Expression of Recombinant Human Insulin Glargine (hIG) in Methylotrophic Yeast Pichia pastoris Herawati, Neng; Rubiyana, Yana; Desriani, Desriani; Santoso, Adi
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.n1.13-20

Abstract

World health organization (WHO) announced that diabetic patients increased significantly yearly worldwide. Consequently, the need for insulin becomes very large. Pichia pastoris, defined as methylotrophic yeast and a well-known expression and protein production host, is widely used for biopharmaceutical-based drug production. This research aims to synthesize human insulin glargine (hIG) protein in yeast P. pastoris. Human insulin glargine is a group of long-acting analogue insulin.We used a synthetic hIG-encoding gene constructed in frame with the truncated α-factor secretion signal in a pD902 expression vector. Recombinant plasmid pD902-hIG was linearized using Sac1 enzyme and transformed into P. pastoris genome. Multicopy clones were selected on YPDS plates containing Zeocin™ with concentrations of 100; 500; 1,000; and 2,000 µg//mL. Analyses using SDS-PAGE, slot blot, and Western blot showed that recombinant hIG protein had been obtained with a molecular weight of approximately 6,000 Daltons.
A Preliminary Report on The Syntheses of Oligonucleotide Primers in The National Research and Innovation Agency (NRIA) Atikana, Akhirta; Prasetyoputri, Anggia; Rubiyana, Yana; Herawati, Neng; Desriani, Desriani; Pratiwi, Riyona Desvy; Wulandari, Dwi; Sukmarini, Linda; Kusharyoto, Wien; Santoso, Adi; Putra, Masteria Yunovilsa; Lisdiyanti, Puspita
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.n1.21-27

Abstract

A PolyGen DNA Synthesizer is equipment that is used for synthesizing oligonucleotide primers for any amplification targets. Oligonucleotide primers are indispensable components for any Polymerase Chain Reaction (PCR)-based detections. In the present study a number of oligonucleotide primer sets were synthesized to target (1) the Human Insuline Glargin (HIG) and (2) the Human Erythropoietin (EPO), as well as (3) the RNA-dependent RNA Polymerase (RdRp) and (4) the Nucleocapsid (N) genes of the severe acute respiratory syndrome virus 2 (SARS-CoV-2). A solid-phase oligonucleotide synthesis method was used according to the default protocol of the Polygen’s instrument to synthesize primers at a 40 nmol scale. The synthesized primers in this study were compared to commercially produced primers in their ability to amplify the gene target(s) in PCR and quantitative real-time PCR (qPCR) reactions. The first two sets of primers showed similar results in PCR compared to commercial primers; however, these primers were not tested for qPCR due to sample limitation. In contrast, the primer sets 3 and 4 were not able to produce amplicons in PCR reactions and only the primer set 4 successfully amplified the gene target in qPCR. These results indicate that the crude primers synthesized in this study are promising candidates for molecular detection and diagnostics, but these primers would benefit from further optimization for routine applications.
Co-Authors Adi Santoso Agustiyanti, Dian Fitria Anam, M. Khairul Andri Wardiana, Andri Atikana, Akhirta Azzahra, Salsabila Desriani Desriani Dwi Wulandari Endah Puji Septisetyani, Endah Puji ERNAWATI ARIFIN GIRI-RACHMAN Ferry Iskandar Fibriani, Azzania Hariyatun, Hariyatun Himawan, Alayna Lillahida Indri Husnaa, Ulfatul Irmanida Batubara Komang Alit Paramitasari Kusharyoto, Wien Kusumawati, Arizah LINDA SUKMARINI Marissa Angelina Neng Herawati, Neng Permatasari, Fitri Aulia Popi Hadi Wisnuwardhani, Popi Hadi Prasetyaningrum, Pekik Wiji Prasetyoputri, Anggia PUSPITA LISDIYANTI Putra, Masteria Yunovilsa Ramadani, Ratna Dwi RATIH ASMANA NINGRUM Riyona Desvy Pratiwi, Riyona Desvy Stephanie, Rebecca Suyoko, Ahmad Taharuddin, Audrey Angelina Putri Wijaya, Sri Kartika Yamahoki, Nicholas