Optimization of a mouse recombinant antibody fragment for efficient production from Escherichia coli

Matrix metalloproteinase 9 (MMP9) is involved in several aspects of the pathology of cancer, including invasion, metastasis, and angiogenesis. In this study, we expressed a recombinant scFv-type anti-MMP9 antibody in soluble form using Escherichia coli, purified it, and confirmed its antigen-binding ability. The convenient, rapid, inexpressive system used in this study for producing recombinant antibody fragments needs only five days, and thus can be used for the efficient production of scFv against MMP9, which can be used in a range of applications and industrial fields, including diagnosis and treatment of inflammatory and cancer-related diseases.

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Use of a design of experiments approach to optimise production of a recombinant antibody fragment in the periplasm of Escherichia coli: selection of signal peptide and optimal growth conditions

AMB Express ◽

10.1186/s13568-018-0727-8 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 7

Author(s):

Ikhlaas M. Kasli ◽

Owen R. T. Thomas ◽

Tim W. Overton

Keyword(s):

Escherichia Coli ◽

Design Of Experiments ◽

Signal Peptide ◽

Recombinant Antibody ◽

Antibody Fragment ◽

Optimal Growth ◽

Growth Conditions ◽

Recombinant Antibody Fragment ◽

Selection Of

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Expression of Recombinant Antibody Fragment, Anti BNP-SCFV on the Periplasm of Escherichia Coli for the Detection of Heart Failure

Molekul ◽

10.20884/1.jm.2017.12.1.288 ◽

2017 ◽

Vol 12 (1) ◽

pp. 30

Author(s):

Shabarni Gaffar ◽

Sofyan Multazam N Aji ◽

Yeni W Hartati ◽

Safri Ishmayana ◽

Toto Subroto

Keyword(s):

Heart Failure ◽

Escherichia Coli ◽

Recombinant Antibody ◽

Antibody Fragment ◽

Single Chain ◽

Recombinant Antibody Fragment ◽

Sds Page ◽

Patients With Heart Failure ◽

Heart Muscle Cells ◽

Antigen Antibody

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.

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