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Parallel production and verification of protein products using a novel high-throughput screening method.
Tegel, Hanna; Yderland, Louise; Boström, Tove; Eriksson, Cecilia; Ukkonen, Kaisa; Vasala, Antti; Neubauer, Peter; Ottosson, Jenny; Hober, Sophia.
Afiliação
  • Tegel H; School of Biotechnology, Royal Institute of Technology, AlbaNova University Center, Stockholm, Sweden.
Biotechnol J ; 6(8): 1018-25, 2011 Aug.
Article em En | MEDLINE | ID: mdl-21681961
ABSTRACT
Protein production and analysis in a parallel fashion is today applied in laboratories worldwide and there is a great need to improve the techniques and systems used for this purpose. In order to save time and money, a fast and reliable screening method for analysis of protein production and also verification of the protein product is desired. Here, a micro-scale protocol for the parallel production and screening of 96 proteins in plate format is described. Protein capture was achieved using immobilized metal affinity chromatography and the product was verified using matrix-assisted laser desorption ionization time-of-flight MS. In order to obtain sufficiently high cell densities and product yield in the small-volume cultivations, the EnBase® cultivation technology was applied, which enables cultivation in as small volumes as 150 µL. Here, the efficiency of the method is demonstrated by producing 96 human, recombinant proteins, both in micro-scale and using a standard full-scale protocol and comparing the results in regard to both protein identity and sample purity. The results obtained are highly comparable to those acquired through employing standard full-scale purification protocols, thus validating this method as a successful initial screening step before protein production at a larger scale.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes / Ensaios de Triagem em Larga Escala Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2011 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes / Ensaios de Triagem em Larga Escala Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2011 Tipo de documento: Article