Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 1 de 1
Filtrar
Mais filtros








Base de dados
Assunto principal
Intervalo de ano de publicação
1.
Nat Commun ; 11(1): 1816, 2020 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-32286330

RESUMO

Protein biopharmaceuticals are highly successful, but their utility is compromised by their propensity to aggregate during manufacture and storage. As aggregation can be triggered by non-native states, whose population is not necessarily related to thermodynamic stability, prediction of poorly-behaving biologics is difficult, and searching for sequences with desired properties is labour-intensive and time-consuming. Here we show that an assay in the periplasm of E. coli linking aggregation directly to antibiotic resistance acts as a sensor for the innate (un-accelerated) aggregation of antibody fragments. Using this assay as a directed evolution screen, we demonstrate the generation of aggregation resistant scFv sequences when reformatted as IgGs. This powerful tool can thus screen and evolve 'manufacturable' biopharmaceuticals early in industrial development. By comparing the mutational profiles of three different immunoglobulin scaffolds, we show the applicability of this method to investigate protein aggregation mechanisms important to both industrial manufacture and amyloid disease.


Assuntos
Agregados Proteicos , Sequência de Aminoácidos , Substituição de Aminoácidos , Regiões Determinantes de Complementaridade/química , Escherichia coli/metabolismo , Humanos , Imunoglobulina G/química , Viabilidade Microbiana , Mutação/genética , Anticorpos de Cadeia Única/química , beta-Lactamases/química
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA