De Novo Isolation & Affinity Maturation of yeast-displayed Virion-binding human fibronectin domains by flow cytometric screening against Virions.
AIDS
Flow cytometry
Human Immunodeficiency Virus
Zika virus
antibody engineering
directed evolution
fibronectin
phage display
protein engineering
yeast display
Journal
Journal of biological engineering
ISSN: 1754-1611
Titre abrégé: J Biol Eng
Pays: England
ID NLM: 101306640
Informations de publication
Date de publication:
2019
2019
Historique:
received:
19
06
2019
accepted:
04
09
2019
entrez:
23
10
2019
pubmed:
23
10
2019
medline:
23
10
2019
Statut:
epublish
Résumé
The promise of biopharmaceuticals comprising one or more binding domains motivates the development of novel methods for de novo isolation and affinity maturation of virion-binding domains. Identifying avenues for overcoming the challenges associated with using virions as screening reagents is paramount given the difficulties associated with obtaining high-purity virus-associated proteins that retain the conformation exhibited on the virion surface. Fluorescence activated cell sorting (FACS) of 1.5 × 10 FACS of a yeast-displayed binding domain library is an efficient method for de novo isolation of virion-binding domains. Affinities of isolated virion-binding clones are readily enhanced via FACS screening of mutant progeny libraries. Given that most binding domains are compatible with yeast display, the approach taken in this work may be broadly utilized for generating virion-binding domains against many different viruses for use in passive immunotherapy and the prevention of viral infection.
Sections du résumé
BACKGROUND
BACKGROUND
The promise of biopharmaceuticals comprising one or more binding domains motivates the development of novel methods for de novo isolation and affinity maturation of virion-binding domains. Identifying avenues for overcoming the challenges associated with using virions as screening reagents is paramount given the difficulties associated with obtaining high-purity virus-associated proteins that retain the conformation exhibited on the virion surface.
RESULTS
RESULTS
Fluorescence activated cell sorting (FACS) of 1.5 × 10
CONCLUSIONS
CONCLUSIONS
FACS of a yeast-displayed binding domain library is an efficient method for de novo isolation of virion-binding domains. Affinities of isolated virion-binding clones are readily enhanced via FACS screening of mutant progeny libraries. Given that most binding domains are compatible with yeast display, the approach taken in this work may be broadly utilized for generating virion-binding domains against many different viruses for use in passive immunotherapy and the prevention of viral infection.
Identifiants
pubmed: 31636701
doi: 10.1186/s13036-019-0203-2
pii: 203
pmc: PMC6796422
doi:
Types de publication
Journal Article
Langues
eng
Pagination
76Subventions
Organisme : NIAID NIH HHS
ID : DP2 AI136600
Pays : United States
Informations de copyright
© The Author(s). 2019.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare that they have no competing interests.
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