Microbial approach of epitope tagged MFE-23 single fragment antibodies production
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 w...
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| Format: | Article |
| Language: | English |
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Faculty of Medicine
2020-12-01
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| Series: | JKKI (Jurnal Kedokteran dan Kesehatan Indonesia) |
| Subjects: | |
| Online Access: | https://journal.uii.ac.id/JKKI/article/view/11674 |
| _version_ | 1797785408470253568 |
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| author | Razmaeda Sarastry |
| author_facet | Razmaeda Sarastry |
| author_sort | Razmaeda Sarastry |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T00:53:41Z |
| format | Article |
| id | doaj.art-e75edd3591784e14a8ae8994704463be |
| institution | Directory Open Access Journal |
| issn | 2085-4145 2527-2950 |
| language | English |
| last_indexed | 2024-03-13T00:53:41Z |
| publishDate | 2020-12-01 |
| publisher | Faculty of Medicine |
| record_format | Article |
| series | JKKI (Jurnal Kedokteran dan Kesehatan Indonesia) |
| spelling | doaj.art-e75edd3591784e14a8ae8994704463be2023-07-07T07:41:36ZengFaculty of MedicineJKKI (Jurnal Kedokteran dan Kesehatan Indonesia)2085-41452527-29502020-12-0122423110.20885/JKKI.Vol11.Iss3.art39707Microbial approach of epitope tagged MFE-23 single fragment antibodies productionRazmaeda Sarastry0Mutiara Bunda Maternal and Child Hospital Salatiga, SalatigaBackground: 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.https://journal.uii.ac.id/JKKI/article/view/11674scfv-mfeceae. colielisa. |
| spellingShingle | Razmaeda Sarastry Microbial approach of epitope tagged MFE-23 single fragment antibodies production JKKI (Jurnal Kedokteran dan Kesehatan Indonesia) scfv-mfe cea e. coli elisa. |
| title | Microbial approach of epitope tagged MFE-23 single fragment antibodies production |
| title_full | Microbial approach of epitope tagged MFE-23 single fragment antibodies production |
| title_fullStr | Microbial approach of epitope tagged MFE-23 single fragment antibodies production |
| title_full_unstemmed | Microbial approach of epitope tagged MFE-23 single fragment antibodies production |
| title_short | Microbial approach of epitope tagged MFE-23 single fragment antibodies production |
| title_sort | microbial approach of epitope tagged mfe 23 single fragment antibodies production |
| topic | scfv-mfe cea e. coli elisa. |
| url | https://journal.uii.ac.id/JKKI/article/view/11674 |
| work_keys_str_mv | AT razmaedasarastry microbialapproachofepitopetaggedmfe23singlefragmentantibodiesproduction |