Identification of recombinant Fabs for structural and functional characterization of HIV-host factor complexes Article Swipe
Natalia Sevillano
,
Evan M. Green
,
Jörg Votteler
,
Dong Young Kim
,
Xuefeng Ren
,
Bei Yang
,
Xi Liu
,
André Luiz Lourenço
,
James H. Hurley
,
Shauna Farr-Jones
,
John D. Gross
,
Yifan Cheng
,
Charles S. Craik
·
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pone.0250318
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pone.0250318
Viral infection and pathogenesis is mediated by host protein—viral protein complexes that are important targets for therapeutic intervention as they are potentially less prone to development of drug resistance. We have identified human, recombinant antibodies (Fabs) from a phage display library that bind to three HIV-host complexes. We used these Fabs to 1) stabilize the complexes for structural studies; and 2) facilitate characterization of the function of these complexes. Specifically, we generated recombinant Fabs to Vif-CBF-β-ELOB-ELOC (VCBC); ESCRT-I complex and AP2-complex. For each complex we measured binding affinities with K D values of Fabs ranging from 12–419 nM and performed negative stain electron microscopy (nsEM) to obtain low-resolution structures of the HIV-Fab complexes. Select Fabs were converted to scFvs to allow them to fold intracellularly and perturb HIV-host protein complex assembly without affecting other pathways. To identify these recombinant Fabs, we developed a rapid screening pipeline that uses quantitative ELISAs and nsEM to establish whether the Fabs have overlapping or independent epitopes. This pipeline approach is generally applicable to other particularly challenging antigens that are refractory to immunization strategies for antibody generation including multi-protein complexes providing specific, reproducible, and renewable antibody reagents for research and clinical applications. The curated antibodies described here are available to the scientific community for further structural and functional studies on these critical HIV host-factor proteins.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1371/journal.pone.0250318
- https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0250318&type=printable
- OA Status
- gold
- References
- 31
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W3163941859
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W3163941859Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1371/journal.pone.0250318Digital Object Identifier
- Title
-
Identification of recombinant Fabs for structural and functional characterization of HIV-host factor complexesWork title
- Type
-
articleOpenAlex work type
- Language
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enPrimary language
- Publication year
-
2021Year of publication
- Publication date
-
2021-05-13Full publication date if available
- Authors
-
Natalia Sevillano, Evan M. Green, Jörg Votteler, Dong Young Kim, Xuefeng Ren, Bei Yang, Xi Liu, André Luiz Lourenço, James H. Hurley, Shauna Farr-Jones, John D. Gross, Yifan Cheng, Charles S. CraikList of authors in order
- Landing page
-
https://doi.org/10.1371/journal.pone.0250318Publisher landing page
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https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0250318&type=printableDirect link to full text PDF
- Open access
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YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
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-
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0250318&type=printableDirect OA link when available
- Concepts
-
Recombinant DNA, Phage display, Biology, Antibody, Computational biology, Epitope, Virology, Biochemistry, Genetics, GeneTop concepts (fields/topics) attached by OpenAlex
- Cited by
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0Total citation count in OpenAlex
- References (count)
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31Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.critical | 216 |
| abstract_inverted_index.electron | 102 |
| abstract_inverted_index.function | 65 |
| abstract_inverted_index.identify | 136 |
| abstract_inverted_index.measured | 85 |
| abstract_inverted_index.mediated | 5 |
| abstract_inverted_index.negative | 100 |
| abstract_inverted_index.pipeline | 145, 163 |
| abstract_inverted_index.reagents | 191 |
| abstract_inverted_index.research | 193 |
| abstract_inverted_index.studies; | 58 |
| abstract_inverted_index.affecting | 132 |
| abstract_inverted_index.available | 203 |
| abstract_inverted_index.community | 207 |
| abstract_inverted_index.complexes | 10, 55, 184 |
| abstract_inverted_index.converted | 116 |
| abstract_inverted_index.described | 200 |
| abstract_inverted_index.developed | 141 |
| abstract_inverted_index.epitopes. | 161 |
| abstract_inverted_index.establish | 153 |
| abstract_inverted_index.generally | 166 |
| abstract_inverted_index.generated | 71 |
| abstract_inverted_index.important | 13 |
| abstract_inverted_index.including | 182 |
| abstract_inverted_index.infection | 1 |
| abstract_inverted_index.pathways. | 134 |
| abstract_inverted_index.performed | 99 |
| abstract_inverted_index.proteins. | 219 |
| abstract_inverted_index.providing | 185 |
| abstract_inverted_index.renewable | 189 |
| abstract_inverted_index.screening | 144 |
| abstract_inverted_index.specific, | 186 |
| abstract_inverted_index.stabilize | 53 |
| abstract_inverted_index.affinities | 87 |
| abstract_inverted_index.antibodies | 34, 199 |
| abstract_inverted_index.applicable | 167 |
| abstract_inverted_index.complexes. | 46, 68, 112 |
| abstract_inverted_index.facilitate | 61 |
| abstract_inverted_index.functional | 212 |
| abstract_inverted_index.generation | 181 |
| abstract_inverted_index.identified | 31 |
| abstract_inverted_index.microscopy | 103 |
| abstract_inverted_index.refractory | 175 |
| abstract_inverted_index.scientific | 206 |
| abstract_inverted_index.strategies | 178 |
| abstract_inverted_index.structural | 57, 210 |
| abstract_inverted_index.structures | 108 |
| abstract_inverted_index.challenging | 171 |
| abstract_inverted_index.development | 25 |
| abstract_inverted_index.host-factor | 218 |
| abstract_inverted_index.independent | 160 |
| abstract_inverted_index.overlapping | 158 |
| abstract_inverted_index.potentially | 21 |
| abstract_inverted_index.recombinant | 33, 72, 138 |
| abstract_inverted_index.resistance. | 28 |
| abstract_inverted_index.therapeutic | 16 |
| abstract_inverted_index.AP2-complex. | 80 |
| abstract_inverted_index.immunization | 177 |
| abstract_inverted_index.intervention | 17 |
| abstract_inverted_index.particularly | 170 |
| abstract_inverted_index.pathogenesis | 3 |
| abstract_inverted_index.quantitative | 148 |
| abstract_inverted_index.Specifically, | 69 |
| abstract_inverted_index.applications. | 196 |
| abstract_inverted_index.multi-protein | 183 |
| abstract_inverted_index.reproducible, | 187 |
| abstract_inverted_index.low-resolution | 107 |
| abstract_inverted_index.intracellularly | 124 |
| abstract_inverted_index.protein—viral | 8 |
| abstract_inverted_index.characterization | 62 |
| abstract_inverted_index.Vif-CBF-β-ELOB-ELOC | 75 |
| cited_by_percentile_year | |
| corresponding_author_ids | https://openalex.org/A5087001996 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 13 |
| corresponding_institution_ids | https://openalex.org/I180670191 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/3 |
| sustainable_development_goals[0].score | 0.7699999809265137 |
| sustainable_development_goals[0].display_name | Good health and well-being |
| citation_normalized_percentile.value | 0.07742797 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |