Protein Purification

A special issue of Biomedicines (ISSN 2227-9059). This special issue belongs to the section "Molecular and Translational Medicine".

Deadline for manuscript submissions: closed (30 April 2024) | Viewed by 5395

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Guest Editor
Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA
Interests: protein purification; protein activity; protein folding
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Special Issue Information

Dear Colleagues,

This Special Issue, titled "Protein Purification", is dedicated to publishing the latest advancements, techniques, and research in the field of protein purification. This interdisciplinary Special Issue provides a platform for scientists, researchers, and experts to share their insights into the purification of proteins, offering valuable knowledge on various methodologies, tools, and applications. Covering topics from fundamental principles to innovative technologies, "Protein Purification" serves as an essential resource for professionals engaged in biochemistry, biotechnology, and other related disciplines, fostering collaboration and progress in this critical area of scientific inquiry. Protein purification is a vital process that underpins our ability to decipher protein structures and advance the development of targeted drugs. It plays a pivotal role in the production of vaccines and pharmaceuticals, making it indispensable in the quest for effective treatments and medical solutions.

Dr. Ibrar Siddique
Guest Editor

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Keywords

  • protein
  • purity
  • yield
  • activity
  • expression

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Published Papers (1 paper)

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19 pages, 2954 KiB  
Protocol
Optimizing Recombinant Cas9 Expression: Insights from E. coli BL21(DE3) Strains for Enhanced Protein Purification and Genome Editing
by Shilpi Agrawal, Made Harumi Padmaswari, Abbey L. Stokes, Daniel Maxenberger, Morgan Reese, Adila Khalil and Christopher E. Nelson
Biomedicines 2024, 12(6), 1226; https://doi.org/10.3390/biomedicines12061226 - 31 May 2024
Cited by 1 | Viewed by 5027
Abstract
The CRISPR-Cas9 system is a revolutionary tool in genetic engineering, offering unprecedented precision and efficiency in genome editing. Cas9, an enzyme derived from bacteria, is guided by RNA to edit DNA sequences within cells precisely. However, while CRISPR-Cas9 presents notable benefits and encouraging [...] Read more.
The CRISPR-Cas9 system is a revolutionary tool in genetic engineering, offering unprecedented precision and efficiency in genome editing. Cas9, an enzyme derived from bacteria, is guided by RNA to edit DNA sequences within cells precisely. However, while CRISPR-Cas9 presents notable benefits and encouraging outcomes as a molecular tool and a potential therapeutic agent, the process of producing and purifying recombinant Cas9 protein remains a formidable hurdle. In this study, we systematically investigated the expression of recombinant SpCas9-His in four distinct Escherichia coli (E. coli) strains (Rosetta2, BL21(DE3), BL21(DE3)-pLysS, and BL21(DE3)-Star). Through optimization of culture conditions, including temperature and post-induction time, the BL21(DE3)-pLysS strain demonstrated efficient SpCas9 protein expression. This study also presents a detailed protocol for the purification of recombinant SpCas9, along with detailed troubleshooting tips. Results indicate successful SpCas9 protein expression using E. coli BL21(DE3)-pLysS at 0.5 mM IPTG concentration. Furthermore, the findings suggest potential avenues for further enhancements, paving the way for large-scale Cas9 production. This research contributes valuable insights into optimizing E. coli strains and culture conditions for enhanced Cas9 expression, offering a step forward in the development of efficient genome editing tools and therapeutic proteins. Full article
(This article belongs to the Special Issue Protein Purification)
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