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Publication
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| Research Title |
Optimization of Transfection Reagents for Transient Expression of Recombinant Antibody in HEK293T Cell |
| Date of Distribution |
21 June 2024 |
| Conference |
| Title of the Conference |
The 4th International Conference on Science Technology & Innovation (4th ICSTI) |
| Organiser |
Maejo University |
| Conference Place |
Chulabhorn Building, Faculty of Science, Maejo University, Chiang Mai, Thailand |
| Province/State |
Chiang Mai, Thailand |
| Conference Date |
29 March 2024 |
| To |
29 March 2024 |
| Proceeding Paper |
| Volume |
2022 |
| Issue |
4 |
| Page |
187-193 |
| Editors/edition/publisher |
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| Abstract |
Recombinant antibody (rAb) has become the crucial platform of monoclonal antibody
production with the fast production, high quality, excellent reproducibility, and its ability to manipulate
antibody formats and properties. Human embryonic kidney 293T (HEK293T) cell is a main expression
host to produce rAbs due to the facilitation of native antibody folding and post-translational modifications.
Generally, rAb expression systems utilize the co-transfection of heavy chain (HC) and light chain (LC)
genes encoded on separate plasmids. The aim of this work is to optimize the transfection reagents to
obtain the high efficient co-transfection method for producing human IgG rAb in HEK293T cell. Here,
we compared the transfection efficiency of two transfection reagents including polyethyleneimine (PEI)
and jetPRIME® (JP). The transfection efficiency of each transfection reagent was tested by human IgG
ELISA for expression and secretion levels of rAb. According to the rAb expression results, jetPRIME®
was identified as the best transfection reagent. Optimal condition of JP was shown at 0.5 μg of each
plasmid DNA (total = 1 μg plasmid DNA), 2 μL of jetPRIME® (DNA:JP ratio = 1:2), and 1.1 × 106 cells of
HEK293T cell. Then, our expressed rAb was purified by affinity chromatography and characterized by
immunofluorescence assay (IFA) for specific binding activities. This study provides detailed protocol of
transfection reagent that allows the effective expression and purification to facilitate small-scale rAb
production by transient co-transfection in a cost- and time-efficient manner. |
| Author |
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| Peer Review Status |
มีผู้ประเมินอิสระ |
| Level of Conference |
นานาชาติ |
| Type of Proceeding |
Full paper |
| Type of Presentation |
Poster |
| Part of thesis |
true |
| Presentation awarding |
false |
| Attach file |
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| Citation |
0
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Journal Publication
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