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All Journal Chimica et Natura Acta Molekul: Jurnal Ilmiah Kimia
Shabarni Gaffar
Departemen Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Padjadjaran
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Environmental Science Materials Science & Nanotechnology

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DETEKSI URUTAN OLIGONUKLEOTIDA Mycobacterium tuberculosis SECARA VOLTAMMETRI MENGGUNAKAN SCREEN PRINTED CARBON ELECTRODE (SPCE) Yeni Wahyuni Hartati; Yohan Yohan; Ratna Nurmalasari; Shabarni Gaffar; Rubianto A. Lubis
Chimica et Natura Acta Vol 4, No 2 (2016)
Publisher : Departemen Kimia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (204.19 KB) | DOI: 10.24198/cna.v4.n2.10674

Abstract

Mycobacterium tuberculosis merupakan bakteri penyebab tuberkulosis (TB). Pengembangan analisis secara biosensor DNA sangat menarik perhatian karena penerapannya mudah. Dalam penelitian ini telah dilakukan penentuan urutan pendek oligonukleotida M. tuberculosis gen RV0508 dari strain H37RV secara voltammetri pulsa differensial menggunakan screen printed carbon electrode (SPCE). Pendeteksian berdasarkan teknik hibridisasi urutan oligonukleotida probe yang diadsorpsi pada permukaan SPCE, dengan pasangan komplementernya dari DNA target, tanpa indikator hibridisasi, yaitu dengan mensubstitusi basa guanin probe dengan basa inosin. Respon hibridisasi berupa signal guanin DNA target di daerah potensial sekitar +0,9 V. Telah diperoleh korelasi linear antara arus puncak dengan konsentrasi DNA target pada rentang 5,0-20,0 µg/mL dengan batas deteksi 8,2 µg/mL.
Expression of Recombinant Antibody Fragment, Anti BNP-SCFV on the Periplasm of Escherichia Coli for the Detection of Heart Failure Shabarni Gaffar; Sofyan Multazam N Aji; Yeni W Hartati; Safri Ishmayana; Toto Subroto
Molekul Vol 12, No 1 (2017)
Publisher : Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (448.733 KB) | DOI: 10.20884/1.jm.2017.12.1.288

Abstract

Basic natriuretic peptide (BNP) is a polypeptide hormone consist of 32 amino acids that secreted by the heart ventricle to respond the excessive stretching of heart muscle cells. BNP can be used as prognostic marker for patients with heart failure. The presence of BNP in blood can be detected by BNP antibody, which is anti BNP-single chain variable fragment (Anti BNP-SCFV). The antibody is a combination of polypeptides between varying region on the heavy chain (VH) and the light chain (VL) of immunoglobulin. Anti BNP-SCFV will bind to BNP through the antigen-antibody interaction. Concentration of BNP in a patient’s blood can be detected through the interaction of BNP with Anti BNP-SCFV using immunosensor method. Production of recombinant Anti BNP-SCFV in Escherichia coli as host is reported in the present study. Anti BNP-SCFV was expressed in fusion form with OmpC signal peptide that direct the protein to a periplasmic space. Expression was performed under RhaBad promoter as control using L-rhamnose as inducer. SDS-PAGE characterization showed consistent band at 28 kDa, which was assumed as Anti BNP-SCFV. The optimum expression was found at four hours after induction with 4 mM inducer. Anti BNP-SCFV was secreted from the cell as characterized by the presence of the protein on periplasmic membrane and extracellular fraction.
Co-Authors Nurmalasari, Ratna Rubianto A. Lubis Safri Ishmayana Sofyan Multazam N Aji Toto Subroto Yeni W Hartati Yeni Wahyuni Hartati Yohan Yohan