Chapter title |
A Screening Methodology for Purifying Proteins with Aggregation Problems
|
---|---|
Chapter number | 14 |
Book title |
Insoluble Proteins
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2205-5_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2204-8, 978-1-4939-2205-5
|
Authors |
Mario Lebendiker, Michal Maes, Assaf Friedler, Lebendiker M, Maes M, Friedler A, Lebendiker, Mario, Maes, Michal, Friedler, Assaf |
Abstract |
Many proteins are prone to aggregate or insoluble for different reasons. This poses an extraordinary challenge at the expression level, but even more during downstream purification processes. Here we describe a strategy that we developed for purifying prone-to-aggregate proteins. Our methodology can be easily implemented in small laboratories without the need for automated, expensive platforms. This procedure is especially suitable for intrinsically disordered proteins (IDPs) and for proteins with intrinsically disordered regions (IDRs). Such proteins are likely to aggregate due to their lack of tertiary structure and their extended and flexible conformations. Similar methodologies can be applied to other proteins with comparable tendency to aggregate during the expression or purification steps.In this chapter, we will mainly focus on protein solubility and stability issues during purification and storage, on factors that can prevent aggregation or maintain solubility, and on the importance of the early elimination of aggregates during protein purification. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 56 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 13 | 23% |
Researcher | 12 | 21% |
Student > Bachelor | 8 | 14% |
Student > Master | 8 | 14% |
Professor | 2 | 4% |
Other | 3 | 5% |
Unknown | 10 | 18% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 17 | 30% |
Agricultural and Biological Sciences | 13 | 23% |
Chemistry | 8 | 14% |
Nursing and Health Professions | 1 | 2% |
Psychology | 1 | 2% |
Other | 3 | 5% |
Unknown | 13 | 23% |