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All Journal Jurnal Kimia Terapan Indonesia Molekul: Jurnal Ilmiah Kimia
Shabarni Gaffar
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Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Materials Science & Nanotechnology

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An Electrochemical Immunosensor for the Detection of B-Type Natriuretic Peptide (BNP) Based on Sandwich ELISA Using Screen Printed Carbon Electrodes Yeni Wahyuni Hartati; Muhammad Hilman Daniswara; Ratna Nurmalasari; Shabarni Gaffar; Toto Subroto
Molekul Vol 13, No 1 (2018)
Publisher : Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (448.537 KB) | DOI: 10.20884/1.jm.2018.13.1.379

Abstract

B-type natriuretic peptide (BNP) is a 32-amino acid polypeptide, a cardiac neuro hormone that specifically secreted from heart ventricle as a response towards the increase of volume and pressure in the heart. The determination of BNP concentration in patients blood is one of the method used to diagnose heart failure. An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes for the detection of the BNP antigen was developed in this study. Monoclonal anti-BNP capture antibody was immobilized on streptavidin-modified SPCEs to give a well oriented of antibody. Furthermore, a biotinylated anti-BNP that conjugated with horseradish peroxidase (HRP) was used as secondary antibody. The electrochemical signal produced by redox activity of substrate 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H2O2) was measured by differential pulse voltammetry. The BNP immunosensor showed a linear response between 1.0×10-2 and 1.0×102 ng/mL, and the limit of detection was 3.3 ng/mL. BNP immunosensor is a promising technology for the rapid and convenient detection of heart failure.
PENGARUH KONSENTRASI PENGINDUKSI METANOL SERTA"SUMBER KARBON SORBITOL DAN MONITOL TERHADAP PRODUKSI a-AMILASE Saccharomycopsis fibuligera R64 DALAM Pichia pastoris Shabarni Gaffar; Dani Permana; Diana P Rahmawati; Triana N Meirina; Abu Bakar M.I. Syihab; Safrl Ismayana; Toto Subroto; O Suprijana
Jurnal Kimia Terapan Indonesia Vol 13, No 2 (2011)
Publisher : Research Center for Chemistry - LIPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (5297.927 KB) | DOI: 10.14203/jkti.v13i2.152

Abstract

Pichia pastoris has been widely used as host for heterologous protein expression for commercial purposes. The advantages of P. pastoris as protein expression host is its ability to grow with high cell density and the presence of a gene promoter that canbe induced tightly, named AOXI encoding alcohol oxidase. Improvement of recombinant protein production under control of AOXI promoter in P. pastoris expression system is still a major concern. Optimization of methanol concentration as inducerand addition of carbon sources, is one of the strategies to improve the expression level. This research aims to study the effect of methanol as inducer as well as sorbitol and mannitol as additional carbon source to the expression level of recombinantSaccharomycopsis fibuligera a-amylase (Sfamy) by P. pastoris (Mut). Sorbitol and manuot known as non-repressive carbon source, to increase the growth ofP. past oris, but not inhibit the AOX1 promoter and foreign proteins expression. Sfamy was expressed in P. pastoris GS115 (His-, Mut) with addition ofcarbon source, sorbitol and mannitol saparately to the expression medium. The result showed that, the optimum concentration" of methanol inducer for Sfamy production is 0.75%. The addition of sorbitol or mannitol increased Sfamy production. Concentrations of sorbitol and mannitol 2% with0.75% methanol inducer increase the secretion level of recombinant Sfamy 2.13 times and 1.94 times respectively, compared with no additional carbon source. This result indicated that addition of carbon source can improved recombinant protein production by P. pastoris, and the used of sorbitol as additionalcarbon source is more effective compared to mannitol.Key words: a-amylase. Pichid pastoris, methanol, sorbitol,mannitol.
Co-Authors Abu Bakar M.I. Syihab Dani Permana Diana P Rahmawati Muhammad Hilman Daniswara Nurmalasari, Ratna O Suprijana Safrl Ismayana Toto Subroto Triana N Meirina Yeni Wahyuni Hartati