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All Journal HAYATI Journal of Biosciences VALENSI Bionatura Microbiology Indonesia Majalah Kedokteran Bandung Jurnal Riset Kimia Indonesian Journal of Pharmaceutical Science and Technology JC-T (Journal Cis-Trans): Jurnal Kimia dan Terapannya Health Notions Chimica et Natura Acta ALCHEMY Jurnal Penelitian Kimia Jurnal Kimia Terapan Indonesia Jurnal Pendidikan Kimia (JPKim) Indonesian Journal of Chemistry PENDIPA Journal of Science Education Molekul: Jurnal Ilmiah Kimia Jurnal Ilmiah Farmako Bahari

Shabarni Gaffar

Departemen Kimia, Fakultas Matematika Dan Ilmu Pengetahuan Alam, Universitas Padjadjaran

Author-ID : 433762

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Dentistry Earth & Planetary Sciences Education Energy Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Nursing Public Health

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Combination of Genetic Manipulation Improved Saccharomycopsis fibuligera α-Amylase Secretion by Pichia pastoris Shabarni Gaffar; Dessy Natalia; Toto Subroto; Oo Suprijana; Soetijoso Soemitro
Indonesian Journal of Chemistry Vol 19, No 2 (2019)
Publisher : Universitas Gadjah Mada

This study assessed the combinations of genetic manipulation; signal peptide modification, gene dosage increment and co-expression of folding component, to increase Saccharomycopsis fibuligera R64 α-amylase (Sfamy) secretion in Pichia pastoris. Sfamy native signal peptide was replaced with modified signal peptide which contained 15 amino acid of mouse salivary α-amylase signal peptide fused to the pro-region of the signal peptide of Saccharomyces cerevisiae α-mating factor (α-MF). Increase in gene dosage was identified by screening for P. pastoris harboring multicopies of the Sfamy gene. Whereas, co-expression of folding component was done by addition of Protein Disulfide Isomerase (PDI). Expression plasmids harboring Sfamy containing modified signal sequence (pPICZA-MS-Sfamy) was used to transform P. pastoris GS115, and gene dosage increment was screened using zeocin. Effect of PDI co-expression on secretion levels of Sfamy was assessed by constructing the pPIC3.5K-Pdi1 plasmid and introducing into P. pastoris harboring multicopies of MS-Sfamy for expression of Sfamy. Signal peptide modification consequently increased Sfamy secretion by P. pastoris by 3.3-fold compared to native signal peptide. Gene dosage increment had improved Sfamy secretion by 11-fold in P. pastoris [MS-Sfamy] resistant to 2000 μg/mL zeocin, compared to P. pastoris harboring one copy of WT-Sfamy. Hence, PDI co-expression increased the secretion of Sfamy by 2-fold as compared without PDI co-expression. In summary, the combination of genetic manipulation successfully increased Sfamy secretion by 20-fold compared to P. pastoris harboring one copy of WT-Sfamy.

An Electrochemical Aptasensor for the Detection of HER2 as a Breast Cancer Biomarker Based on Gold Nanoparticles-Aptamer Bioconjugates Yeni Wahyuni Hartati; Sari Syahruni; Shabarni Gaffar; Santhy Wyantuti; Muhammad Yusuf; Toto Subroto
Indonesian Journal of Chemistry Vol 21, No 6 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijc.67124

Inaccurate diagnoses contributes to the high mortality rate of breast cancer. Human epidermal growth factor receptor 2 (HER2) is overexpressed in breast cancer tumors at around 20–30%. This study aims to develop an electrochemical biosensor for HER2 based on a gold nanoparticle-aptamer bioconjugate (AuNP@HER2 aptamer) and investigate the interaction between DNA aptamer and HER2 using computational methods. The bioconjugate was synthesized using maleimide and polyethylene glycol as a linker. The –NH2 group of cysteamine that modified the gold electrode can form a covalent bond with the bioconjugate maleimide. The interaction of the bioconjugated aptamer with HER2 was measured electrochemically based on the [Fe(CN)6]3−/4− redox system. The limit of detection, the linear range of HER2, precision, and accuracy in this study were 1.52 ng mL–1, 0.01 to 15.0 ng mL–1, 0.1298, and 94.06%, respectively. The structure of the DNA aptamer was modeled using mFold, Assemble2, and Chimera, with the interaction between the DNA aptamer and HER2 explored by NPDock. The modeling of the aptamer with HER2 showed that electrostatic interactions dominated the attractive forces. The resulting interaction pattern can be used as a template to improve the binding energy of the aptamer, thus providing insight into the development of aptamer-based biosensors.

Aktivitas Antioksidan dan Sitotoksik Terhadap Sel Kanker HeLa dari Ekstrak Daun Vernonia amygdalina (Asteraceae) Shabarni Gaffar; Muhammad Yudha Nugraha; Ersanda Hafiz; Hesti Lina Wiraswati; Tati Herlina
Chimica et Natura Acta Vol 10, No 1 (2022)
Publisher : Departemen Kimia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24198/cna.v10.n1.36779

Vernonia amygdalina atau dikenal sebagai daun afrika merupakan salah satu tanaman obat yang berpotensi untuk dikembangkan sebagai nutraceutical, seperti antidiabetes, antimalaria dan antikanker. Penelitian ini bertujuan menentukan aktivitas antioksidan dan aktivitas sitotoksik ekstrak daun V. amygdalina terhadap sel kanker serviks HeLa, serta menentukan kelompok senyawa yang terdapat dalam ekstrak daun V. amygdalina dengan aktivitas sitotoksik dan antioksidan tertinggi melalui penapisan fitokimia. Metode yang digunakan pada penelitian ini adalah ekstraksi dan fraksinasi menggunakan pelarut metanol, etil asetat, dan n-hexana. Semua fraksi diuji aktivitas antioksidannya menggunakan metoda DPPH dan aktivitas sitotoksiknya terhadap sel kanker serviks HeLa menggunakan metode MTS, serta penapisan fitokimia untuk menentukan kelompok senyawa alkaloid, terpenoid, steroid, dan flavonoid. Hasil penelitian menunjukkan bahwa ekstrak etil asetat memiliki aktivitas antioksidan paling tinggi, IC50 ekstrak metanol, etil asetat dan n-heksana berturut-turut adalah: 25,27; 17,89; 213,4 ppm. Hasil uji aktivitas sitotoksik menunjukkan bahwa ekstrak metanol, etil asetat dan n-heksana daun V. amygdalina bersifat kurang aktif terhadap sel kanker serviks HeLa dengan IC50 berturut-turut: 519,42; 249,18 dan 367,24 ppm. Ekstrak etil asetat yang mengandung aktivitas antioksidan tertinggi mengandung kelompok senyawa flavonoid, alkaloid, steroid, dan terpenoid, sehingga berpotensi untuk dimanfaatkan sebagai agen kemopreventif berdasarkan aktivitas antioksidannya.

AKTIVITAS SITOTOKSIK FRAKSI ETIL ASETAT DAUN KELOR (MORINGA OLEIFERA) DAN PENGARUHNYA TERHADAP INDUKSI APOPTOSIS PADA SEL KANKER PAYUDARA T47D Riza Apriani; Shabarni Gaffar; Tati Herlina
Jurnal Ilmiah Farmako Bahari Vol 10, No 1 (2019): Jurnal Ilmiah Farmako Bahari
Publisher : Fakultas MIPA Universitas Garut

Tanaman kelor (Moringa oleifera) adalah salah satu tanaman yang telah banyak digunakan dalam industri pengobatan tradisional. Beberapa penelitian telah melaporkan bahwa ekstrak daun M. oleifera memiliki aktivitas antiproliferatif pada beberapa sel kanker, di antaranya terhadap sel kanker hati HepG2, sel kanker paru A549, sel kanker kolon Caco-2 dan sel kanker payudara MDA-MB-231, namun penelitian terhadap sel kanker payudara T47D belum dilakukan. Oleh karena itu, penelitian ini bertujuan untuk mengetahui aktivitas antikanker daun M. oleifera terhadap sel kanker payudara T47D. Ekstrak daun M. oleifera diperoleh dengan cara ekstraksi menggunakan pelarut etanol, kemudian difraksionasi dengan pelarut n-heksana dan etil asetat. Uji sitotoksik yang dilakukan pada penelitian ini menggunakan metode MTT assay. Pengujian terhadap induksi apoptosis dilakukan dengan metode pewarnaan Annexin V-PI dan dianalisis dengan flow cytometry. Kajian molekular juga dilakukan melalui pengamatan protein regulator apoptosis, yaitu Bcl-2 dengan metode imunositokimia. Berdasarkan hasil dari MTT assay, sel T47D yang diberi perlakuan dengan fraksi etil asetat daun M. oleifera menunjukkan aktivitas medium sebagai antikanker dengan nilai IC50 sebesar 243,58 μg/mL. Pengujian terhadap apoptosis menunjukkan bahwa fraksi etil asetat daun M. oleifera dapat menginduksi apoptosis. Hasil pengamatan dengan imunositokimia menunjukkan bahwa pemberian fraksi etil asetat daun M. oleifera mampu menurunkan ekspresi protein Bcl-2. Kata kunci: Apoptosis, Moringa oleifera, kanker payudara, sel T47D

Codon Optimization and Chaperone Assisted Solubilization of Recombinant Human Prethrombin-2 Expressed in Escherichia coli SARONOM SILABAN; IMAN PERMANA MAKSUM; SHABARNI GHAFFAR; KHOMAINI HASAN; SUTARYA ENUS; TOTO SUBROTO; SOETIJOSO SOEMITRO
Microbiology Indonesia Vol. 8 No. 4 (2014): December 2014
Publisher : Indonesian Society for microbiology

Prethrombin-2 (PT2) is a thrombin precursor, which plays a role in the conversion of fibrinogen into fibrin during blood clotting process. Previous study reported that the expression of human prothrombin-2 (rhPT2) in Escherichia coli formed inclusion bodies. The aim of this study was to establish a strategy to express a soluble rhPT2 in E. coli. This study was animed to design and codon optimize human prethrombin-2 gene as well as to optimize the expression condition using four strains of E. coli. The codon adaptation index (CAI) of the unoptimized hpt2 gene was 0.336, with 56.8% GC content. After optimization, the CAI of optimized hpt2 became 1.000 with 53.1% GC content. The optimized gene was successfully cloned into pTWIN1 expression vector. Expression analysis indicated that only E. coli ArcticExpress strain could successfully express a soluble recombinant rhPT2 protein, with only part of rhPT2 being expressed in insoluble form. However, the rest of the E. coli strains used in the experiments failed to express the rhPT2 in soluble form. We are deducing that the success in achieving soluble expression was not only due to the availability of chaperonins Cpn60/Cpn10, which played a crucial role in the protein folding in E. coli ArcticExpress strain, but also due to the codon optimization of hpt2 gene.

A Computational Study of Fab CHK152 with CHK265 as Anti Chikungunya Virus for Rapid Test Components Korry Novitriani; Ade Rizki Ridwan Firdaus; Bacthi Alisjahbana; Shabarni Gaffar; Muhammad Yusuf; Toto Subroto
Health Notions Vol 7, No 5 (2023): May
Publisher : Humanistic Network for Science and Technology (HNST)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33846/hn70502

The symptoms of chikungunya disease are generally similar to the symptoms of other viral diseases, except that chikungunya disease can cause joint disorders and even paralysis. Incorrect initial diagnosis will result in inappropriate treatment of the disease Thus it is necessary to study the identified antibodies that can neutralize and protect the body from alphavirus infections, including chikungunya. There are several antibodies that can neutralize chikungunya including CHK152 which neutralizes virus by inhibiting fusion with stabilizing the surface area of the virus, hindering the exposure of the fusion-loop, likely neutralizing infection by blocking fusion. Otherwise CHK265 inhibits the life cycle of the virus in a way neutralize infection by blocking entry at a post-attachment pre-fusion step. The low energy of binding between antigen-antibodies will stabilize the interaction of both. Therefore, this aims to find the best affinity energy for antibodies to be used in the rapid diagnostic component of anti-chikungunya virus (CHIKV). The binding energies were calculated using FireDock an efficient method for the refinement and rescoring of rigid-body docking solutions, through rearrangement of the interface sidechains and adjustment of the relative orientation of the molecules.The results of this study obtained energy scores from CKH152 and CHK265 are -125.78 and -29.94, indicate that CHK265 antibody affinity energy is higher than CHK152. It can be concluded that the best affinity energy for CHIKV diagnostic component is CHK152.Keywords: energy affinity; CHK152; CHK265; E1E2 CHIKV; molecular docking

Triterpenoids from the Stem Bark of Aglaia cucullata (Meliaceae) and Their Cytotoxic Activity against A549 Lung Cancer Cell Line Desi Harneti; Iqbal Wahyu Mustaqim; Darwati Darwati; Al Arofatus Naini; Purnama Purnama; Erina Hilmayanti; Tri Mayanti; Nurlelasari Nurlelasari; Shabarni Gaffar; Rani Maharani; Kindi Farabi; Unang Supratman; Sofa Fajriah; Mohamad Nurul Azmi; Yoshihito Shiono
Indonesian Journal of Chemistry Vol 23, No 4 (2023)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijc.78748

The Aglaia species, which contains triterpenoids, is the most numerous in the Meliaceae family. The A. cucullata species, of which there are only a few known examples, has received scant research attention. This investigation aims to identify triterpenoids in an n-hexane preparation of A. cucullata stem bark and evaluate their effects against the A549 lung cancer cell line. Five dammarane-type triterpenoids were isolated from the A. cucullata trunk bark, which is (1) (20S)-20-hydroxydammar-24-en-3-one, (2) cabraleone, (3) cabralealactone, (4) eichlerianic acid, and (5) (+)-fouquierol. Their chemical structures were determined using infrared, high-resolution mass spectrometry, and nuclear magnetic resonance, as well as through data comparison of the reported compounds. Compound 1 was priorly separated from the Aglaia genus, compounds 2–4 were first isolated from the A. cucullata species, and compound 5 has been reportedly isolated from the Meliaceae family and the Aglaia genus. All substances were tested for their lethal potential against the A549 lung cancer cell type. A seco structure in the A ring of dammarane-type triterpenoid might play an important part in the lethal activity of component 4, which showed the greatest activity with an IC50 value of 32.17 µM against the A549 lung cancer cell line.

The Stability Study of Electrochemical Aptasensor to Detect SARS-CoV-2 Spike Protein and Its Application for Clinical Samples of Nasopharyngeal Swab Arum Kurnia Sari; Ghina Nur Fadhilah; Irkham Irkham; Muhammad Yusuf; Shabarni Gaffar; Yeni Wahyuni Hartati
Indonesian Journal of Chemistry Vol 23, No 2 (2023)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijc.77887

The stability characteristics associated with the shelf life of a biosensor are rarely investigated, however, they are important factors for real applications. Stability is the variation in the detection signal over a long period of storage. This study aims to determine the effect of storage time on the stability of SARS-CoV-2 receptor binding domain (RBD) spike protein aptamers related to shelf life and the performance of an electrochemical aptasensor on clinical samples. The research method includes a stability study conducted using the accelerated stability method based on the Arrhenius equation at three variations of temperature and storage time. The electrochemical aptasensor's performance was evaluated on clinical samples of 32 nasopharyngeal swabs at biosafety level 3 and its potential on clinical saliva samples. The results indicated that the developed electrochemical aptasensor was stable for ± 15 days with a shelf life of 18, 17 and 16 days, respectively, at 25, 40 and 50 °C. This electrochemical aptasensor has the potential to be a Point of Care (POC) device for the clinical detection of SARS-CoV-2 because it can be tested on clinical samples of nasopharyngeal swabs and the results show its potential application to detect in clinical saliva samples.

Modifikasi Screen Printed Carbon Electrode Menggunakan Cerium dan Optimasi Kondisi Percobaan Biosensor Elektrokimia untuk Deteksi DNA Mitokondria Sus Scrofa Fajriyah, Maulida; Hartati, Yenni Wahyuni; Anggraeni, Anni; Gaffar, Shabarni
JC-T (Journal Cis-Trans): Jurnal Kimia dan Terapannya Vol 6, No 1 (2022)
Publisher : State University of Malang or Universitas Negeri Malang (UM)

Penggunaan cerium dalam aplikasi sensor telah banyak dikembangkan. Dalam penelitian ini, biosensor elektrokimia telah dikembangkan untuk mendeteksi DNA mitokondria Sus scrofa menggunakan screen printed carbon elektrode-cerium (SPCE-Ce). Permukaan elektrode SPCE sebelum dan sesudah dimodifikasi dengan cerium dikarakterisasi menggunakan Scanning Electron Microscopy (SEM) dan voltammetri siklik (CV). Desain urutan nukleotida babi sebagai DNA probe, ditentukan dengan menggunakan program NCBI blast dan T-Coffee. DNA probe diamobilisasikan pada SPCE-Ce melalui interaksi elektrostatik antara permukaan cerium dengan tulang punggung fosfat dari DNA. Selanjutnya proses hibridisasi DNA probe - target dikarakterisasi menggunakan voltammetri diferensiasi pulsa (DPV). Permukaan SPCE dan SPCE-Ce secara SEM menunjukkan perbedaan morfologi yaitu permukaan yang lebih halus dan rata setelah permukaan karbon tertutupi cerium. Voltammogram siklik menunjukkan adanya kenaikan puncak arus redoks dari spesi elektroaktif ferisianida pada SPCE-Ce. Urutan DNA probe adalah 5’-TATTIATACCAATCACTAIC-3’ yang memiliki homologi 100% dengan DNA babi dan homologi 0% dengan DNA ayam dan sapi. Persamaan linier konsentrasi target terhadap arus menghasilkan persamaan I = 0,0303 [target] + 3,947 untuk rentang konsentrasi DNA target 5,0 sampai 30,0 µg/mL, dan nilai LoD adalah 1,44 µg/mL. Biosensor ini selanjutnya dapat digunakan untuk penentuan konsentrasi mtDNA babi dalam sampel.

Dysobinol Extracted from Chisocheton Macrophyllus Triggers Proliferation Inhibition, Potential Apoptosis, and Cell Cycle Arrest of He La Cancer Cell Lines Gaffar, Shabarni; Hafiz, Ersanda; Wiraswati, Hesti Lina; Ishmayana, Safri; Nurlelasari, Nurlelasari
Majalah Kedokteran Bandung Vol 56, No 1 (2024)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15395/mkb.v56.3249

Dysobinol is a new limonoid from C. macrophyllus seeds reported to have an anticancer activity. This study aimed to determine the cytotoxic activity of Dysobinol against HeLa cancer cell lines and evaluate its mechanism of action by determining the expression level of several carcinogenesis genes related to apoptosis and cell cycle. In this experimental study, the cytotoxic activity was determined using the MTS assay and gene expression by real-time reverse transcriptase PCR. The result shows that Dysobinol has an anticancer activity in a dose and time-dependent manner against HeLa cells and was categorized as toxic with IC50 values of 52.92, 52.70, and 14.96 μg/ml for 24, 48, and 72 hours, respectively. Dysobinol significantly increased the expression of Bax, Cas-8, and Cas-3 and decreased the expression of Cyc D1 at both doses (IC50 and 2x IC50) but only high doses (2x IC50) could affect Cas9 and NF-κB expressions, indicating that Dysobinol can induce apoptosis via the extrinsic pathway and inhibits the cell cycle through the Cyc D1 regulator. Dysobinol has the potential to be developed as a chemotherapy drug or an adjuvant agent for cervical cancer treatment.

Co-Authors Aathirah, A Sayyidatina Abu Bakar M.I. Syihab Ade Rizki Ridwan Firdaus Aga Adi Masyhuri Agus Safari Al Arofatus Naini Albayyinah, Dyandra Hera Amalia, Riezki Anni Anggraeni, Anni Annisa Ilma Naviardianti Annisa Ilma Naviardianti Ari Hardianto Arum Kurnia Sari Bacthi Alisjahbana Bayu Shiddiq Widhi Pratama Dani Permana Darwati Darwati Desi Harneti Putri Huspa DESSY NATALIA Diana P Rahmawati Egista Istioka Fazrin Ekawardani, Savira Endang Srieatimah - Erina Hilmayanti Ersanda Hafiz Euis Julaeha Euis Julaeha Fajriyah, Maulida Ghina Nur Fadhilah Ghina Uli Felicia Tambunan Hafiz, Ersanda Hesti Lina Wiraswati Iman Permana Maksum Iqbal Wahyu Mustaqim Irkham Irkham KHOMAINI HASAN Kindi Farabi Korry Novitriani, Korry Lia Faridah LINAWATI HARDJITO Lubis, Rubianto A. M. Lutfi Firdaus Ma'ruf, Ilma Fauziah Masyhuri, Aga Adi Mohamad Nurul Azmi Muhammad Hilman Daniswara Muhammad Yudha Nugraha Muhammad Yusuf Murniaty Simorangkir Nayla Haraswati Nurlelasari Nurlelasari Nurmalasari, Ratna Nurul Auliany O Suprijana Oo Suprijana Permadi, Nandang Pertiwi, Wulan Purba Upay Purnama Purnama PUTRI, RAFIKA Putri, Rafika Nanda R. Ukun M.S. Soedjanaatmadja Rani Maharani Rima Melati Rini Surbakti Riza Apriani Riza Apriani Riza Apriani Rubianto A. Lubis Rudi Hartono Rustaman Rustaman Saadah D. Rachman Safri Ishmayana Safrl Ismayana Santhy Wyantuti Sari Syahruni SARONOM SILABAN Saronom Silaban Savira Ekawardhani Shabrinna, Hanif SOETIJOSO SOEMITRO Soetijoso Soemitro Soetijoso Soemitro Sofa Fajriah Sofyan Multazam N Aji Supriyadi, Isma Yustifania SUTARYA ENUS Sutarya Enus Syifa Al Fauziah Rahmani Tambunan, Ghina Uli Felicia Tati Herlina Tati Herlina Toto Subroto Tri Mayanti Triana N Meirina Umi Baroroh Unang Supratman Upay, Purba Vicki Nishinarizki Wulan Pertiwi Yeni W Hartati Yeni Wahyuni Hartati Yenni Wahyuni Hartati, Yenni Wahyuni Yohan Yohan Yohan Yohan Yoshihito Shiono

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