Expression of Recombinant Antibody Fragment, Anti BNP-SCFV on the Periplasm of Escherichia Coli for the Detection of Heart Failure
Journal Title: Molekul - Year 2017, Vol 12, Issue 1
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 patientâs 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.
Authors and Affiliations
Shabarni Gaffar, Sofyan Multazam N Aji, Yeni W Hartati, Safri Ishmayana, Toto Subroto
Keywords
Related Articles
- EP ID EP609176
- DOI 10.20884/1.jm.2017.12.1.288
- Views 89
- Downloads 0
Thermodynamic Properties of Water Adsorption on Gaplek Flour Fortified with Red Bead Tree Seed
The moisture sorption isotherm data of gaplek flour fortified with red bead tree seed stored in a chamber, the relative humidity of chamber ranging from 10% to 97% regulated using a saturated salt solution, was determine...
Effect of Natural Fe2O3 Doping on Performance of Lithium Phospate Ceramic Glass as Secondary Battery Cathode
Natural Fe2O3 doped lithium phosphate Ceramic-glass was made by glass route method at 900 oC with the composition of 5Li2CO3:15ZnO : x Fe2O3: (80 – x) P2O5 where x = 0; 0.5; 1.5 (in mol percent). Thermal stability of s...
Evaluation of Free Radical Scavenging Activity in Ethanolic Extract from Promising Accessions of Curcuma aeruginosa RoxB.
This study evaluated the free radical scavenging activity in ethanolic extracts from 20 accessions of Curcuma aeruginosa. The radical scavenging activity of the extract accessions was investigated with 2,2-diphenyl-1-pic...
Isolation and evaluation of the antioxidant actity of phenolic constituents of the Garcinia sizygiifolia
Garcinia sizygiifolia is a native plant to the South Sulawesi region popularly known as sula and has been cultivated in several regions in Indonesia. The plant by local community use as a food and source of wood, but...
Fractionation And Environmental Risk Of Trace Metals In Surface Sediment Of The East China Sea By Modified BCR Sequential Extraction Method
Geochemical fractionation of Cadmium (Cd), Lead (Pb), Cobalt (Co), Zink (Zn), Iron (Fe) and Manganese (Mn) in sediment of the East China Sea were determined using four-stages of modified BCR sequential extraction method...