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Submitted
November 24, 2018
Accepted
November 25, 2020
Published
December 31, 2020
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Abstract

Background: Antibodies have been investigated for future clinical application in cancer management. An antibody, MFE-23 scFv is known for its ability to bind Carcinoembryonic Antigen (CEA). Different from a full length antibody, single-chain variable fragments (scFvs) are recombinant antibodies in which single polypeptide is engineered to replace variable regions encoding antigen-binding domain. In vitro production of single chain fragment antibodies may use E. coli microorganism for its ability to self-replicating a plasmid.
Objective: This study aimed to produce his- and myc- tagged MFE-23 scFv antibodies by using E. coli culture and to detect their solubility by using ELISA assay.
Methods: Transformed E. coli containing sequences of MFE-23 coding were inoculated and evaluated for their optical density. An ELISA plate was coated by CEA or PBS and secondary antibodies were anti-his, anti-myc and anti-MFE. Horseradish peroxidase-OPD substrate was added to produce chromatic reaction for qualitative detection.
Results: The results showed that each characterized tube was positive for myc-tagged MFE, his- and myc- tagged MFE, and his-tagged MFE for tube 1, 2, and 3 respectively.
Conclusion: This study indicated that transformed E. coli culture is a suitable host for MFE-23 svFV production, and qualitative ELISA assay is a simple useful method for antibody detection and characterization of single chain antibodies.

Keywords

scFv-MFE CEA E. coli ELISA.

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How to Cite
Sarastry, R. (2020). Microbial approach of epitope tagged MFE-23 single fragment antibodies production. JKKI : Jurnal Kedokteran Dan Kesehatan Indonesia, 11(3), 224–231. https://doi.org/10.20885/JKKI.Vol11.Iss3.art3
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