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Quality Screening of Incorrectly Folded Soluble Aggregates from Functional Recombinant Proteins.
Kwon, Soon Bin; Yu, Ji Eun; Kim, Jihoon; Oh, Hana; Park, Chan; Lee, Jinhee; Seong, Baik L.
Afiliación
  • Kwon SB; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. yunbin829@gmail.com.
  • Yu JE; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. 4-season-love@hanmail.net.
  • Kim J; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. wlgns2568@naver.com.
  • Oh H; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. ohanah@naver.com.
  • Park C; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. justies@naver.com.
  • Lee J; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. glory-jjin@hanmail.net.
  • Seong BL; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea. blseong@yonsei.ac.kr.
Int J Mol Sci ; 20(4)2019 Feb 19.
Article en En | MEDLINE | ID: mdl-30791505
ABSTRACT
Solubility is the prime criterion for determining the quality of recombinant proteins, yet it often fails to represent functional activity due to the involvement of non-functional, misfolded, soluble aggregates, which compromise the quality of recombinant proteins. However, guidelines for the quality assessment of soluble proteins have neither been proposed nor rigorously validated experimentally. Using the aggregation-prone enhanced green-fluorescent protein (EGFP) folding reporter system, we evaluated the folding status of recombinant proteins by employing the commonly used sonication and mild lysis of recombinant host cells. We showed that the differential screening of solubility and folding competence is crucial for improving the quality of recombinant proteins without sacrificing their yield. These results highlight the importance of screening out incorrectly folded soluble aggregates at the initial purification step to ensure the functional quality of recombinant proteins.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Pliegue de Proteína / Agregado de Proteínas Tipo de estudio: Diagnostic_studies / Screening_studies Idioma: En Año: 2019 Tipo del documento: Article

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Pliegue de Proteína / Agregado de Proteínas Tipo de estudio: Diagnostic_studies / Screening_studies Idioma: En Año: 2019 Tipo del documento: Article