Direct selection of functional fluorescent-protein antibody fusions by yeast display.
Antibodies are important reagents for research, diagnostics, and therapeutics. Many examples of chimeric proteins combining the specific target recognition of antibodies with complementing functionalities such as fluorescence, toxicity or enzymatic activity have been described. However, antibodies s...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0280930 |
| _version_ | 1811161669679710208 |
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| author | Nileena Velappan Fortunato Ferrara Sara D'Angelo Devin Close Leslie Naranjo Madeline R Bolding Sarah C Mozden Camille B Troup Donna K McCullough Analyssa Gomez Marijo Kedge Andrew R M Bradbury |
| author_facet | Nileena Velappan Fortunato Ferrara Sara D'Angelo Devin Close Leslie Naranjo Madeline R Bolding Sarah C Mozden Camille B Troup Donna K McCullough Analyssa Gomez Marijo Kedge Andrew R M Bradbury |
| author_sort | Nileena Velappan |
| collection | DOAJ |
| description | Antibodies are important reagents for research, diagnostics, and therapeutics. Many examples of chimeric proteins combining the specific target recognition of antibodies with complementing functionalities such as fluorescence, toxicity or enzymatic activity have been described. However, antibodies selected solely on the basis of their binding specificities are not necessarily ideal candidates for the construction of chimeras. Here, we describe a high throughput method based on yeast display to directly select antibodies most suitable for conversion to fluorescent chimera. A library of scFv binders was converted to a fluorescent chimeric form, by cloning thermal green protein into the linker between VH and VL, and directly selecting for both binding and fluorescent functionality. This allowed us to directly identify antibodies functional in the single chain TGP format, that manifest higher protein expression, easier protein purification, and one-step binding assays. |
| first_indexed | 2024-04-10T06:18:04Z |
| format | Article |
| id | doaj.art-11a2f16525564b46ab218f4b5b96ed0c |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-10T06:18:04Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-11a2f16525564b46ab218f4b5b96ed0c2023-03-02T05:31:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01182e028093010.1371/journal.pone.0280930Direct selection of functional fluorescent-protein antibody fusions by yeast display.Nileena VelappanFortunato FerraraSara D'AngeloDevin CloseLeslie NaranjoMadeline R BoldingSarah C MozdenCamille B TroupDonna K McCulloughAnalyssa GomezMarijo KedgeAndrew R M BradburyAntibodies are important reagents for research, diagnostics, and therapeutics. Many examples of chimeric proteins combining the specific target recognition of antibodies with complementing functionalities such as fluorescence, toxicity or enzymatic activity have been described. However, antibodies selected solely on the basis of their binding specificities are not necessarily ideal candidates for the construction of chimeras. Here, we describe a high throughput method based on yeast display to directly select antibodies most suitable for conversion to fluorescent chimera. A library of scFv binders was converted to a fluorescent chimeric form, by cloning thermal green protein into the linker between VH and VL, and directly selecting for both binding and fluorescent functionality. This allowed us to directly identify antibodies functional in the single chain TGP format, that manifest higher protein expression, easier protein purification, and one-step binding assays.https://doi.org/10.1371/journal.pone.0280930 |
| spellingShingle | Nileena Velappan Fortunato Ferrara Sara D'Angelo Devin Close Leslie Naranjo Madeline R Bolding Sarah C Mozden Camille B Troup Donna K McCullough Analyssa Gomez Marijo Kedge Andrew R M Bradbury Direct selection of functional fluorescent-protein antibody fusions by yeast display. PLoS ONE |
| title | Direct selection of functional fluorescent-protein antibody fusions by yeast display. |
| title_full | Direct selection of functional fluorescent-protein antibody fusions by yeast display. |
| title_fullStr | Direct selection of functional fluorescent-protein antibody fusions by yeast display. |
| title_full_unstemmed | Direct selection of functional fluorescent-protein antibody fusions by yeast display. |
| title_short | Direct selection of functional fluorescent-protein antibody fusions by yeast display. |
| title_sort | direct selection of functional fluorescent protein antibody fusions by yeast display |
| url | https://doi.org/10.1371/journal.pone.0280930 |
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