In the last decades, protein engineering has developed particularly in biotechnology and pharmaceutical field. In particular, the engineered antibody subclass has arisen. The single chain diabody format (scDb), conjugating small size with antigen specificity, offers versatility representing a gold standard for a variety of applications, spacing from research to diagnostics and therapy. Along with such advantages, comes the challenge of optimizing their production, improving expression systems, purification procedures and stability. All such parameters are detrimental for protein production in general and above all for recombinant antibody expression, which has to be fine-tuned, choosing a proper protein-expression host and adjusting expression protocols accordingly. In the present paper, we present data regarding the production and purification of a single chain diabody directed against the macromolecular complex hERG1/β1 integrin. We focus on the expression of clones deriving from the transformation of Pichia pastoris yeast cells. In particular, we compare two different clones arose from two separate transformation processes, demonstrating that both are suitable for proper protein expression. Moreover, we have set up an expression protocol and compared the yields obtained using two purification machines: Akta Pure and Akta Start, with a positive outcome.

Expression and purification of a novel single-chain diabody (scDb-hERG1/β1) from Pichia pastoris transformants / Duranti, Claudia; Lastraioli, Elena; Iorio, Jessica; Capitani, Chiara; Carraresi, Laura; Gonnelli, Leonardo; Arcangeli, Annarosa. - In: PROTEIN EXPRESSION AND PURIFICATION. - ISSN 1046-5928. - ELETTRONICO. - 184:(2021), pp. 1-7. [10.1016/j.pep.2021.105879]

Expression and purification of a novel single-chain diabody (scDb-hERG1/β1) from Pichia pastoris transformants

Duranti, Claudia;Lastraioli, Elena;Iorio, Jessica;Capitani, Chiara;Carraresi, Laura;Gonnelli, Leonardo;Arcangeli, Annarosa
2021

Abstract

In the last decades, protein engineering has developed particularly in biotechnology and pharmaceutical field. In particular, the engineered antibody subclass has arisen. The single chain diabody format (scDb), conjugating small size with antigen specificity, offers versatility representing a gold standard for a variety of applications, spacing from research to diagnostics and therapy. Along with such advantages, comes the challenge of optimizing their production, improving expression systems, purification procedures and stability. All such parameters are detrimental for protein production in general and above all for recombinant antibody expression, which has to be fine-tuned, choosing a proper protein-expression host and adjusting expression protocols accordingly. In the present paper, we present data regarding the production and purification of a single chain diabody directed against the macromolecular complex hERG1/β1 integrin. We focus on the expression of clones deriving from the transformation of Pichia pastoris yeast cells. In particular, we compare two different clones arose from two separate transformation processes, demonstrating that both are suitable for proper protein expression. Moreover, we have set up an expression protocol and compared the yields obtained using two purification machines: Akta Pure and Akta Start, with a positive outcome.
2021
184
1
7
Goal 3: Good health and well-being for people
Duranti, Claudia; Lastraioli, Elena; Iorio, Jessica; Capitani, Chiara; Carraresi, Laura; Gonnelli, Leonardo; Arcangeli, Annarosa
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1234960
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