| Autor: |
Kineavy FB; School of Life Sciences, University of Liverpool., Davies AA; School of Life Sciences, University of Liverpool., Mitchell MR; School of Life Sciences, University of Liverpool., Lay D; School of Life Sciences, University of Liverpool., Dominguez MJ; Health Sciences Center, Texas Tech University., Stollar EJ; School of Life Sciences, University of Liverpool; E.Stollar@liverpool.ac.uk. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2021 May 21 (171). Date of Electronic Publication: 2021 May 21. |
| DOI: |
10.3791/62075 |
| Abstrakt: |
Protein purification is imperative to the study of protein structure and function and is usually used in combination with biophysical techniques. It is also a key component in the development of new therapeutics. The evolving era of functional proteomics is fueling the demand for high-throughput protein purification and improved techniques to facilitate this. It was hypothesized that a multi column plate adaptor (MCPA) can interface multiple chromatography columns of different resins with multi-well plates for parallel purification. This method offers an economical and versatile method of protein purification that can be used under gravity or vacuum, rivaling the speed of an automated system. The MCPA can be used to recover milligram yields of protein by an affordable and time efficient method for subsequent characterization and analysis. The MCPA has been used for high-throughput affinity purification of SH3 domains. Ion exchange has also been demonstrated via the MCPA to purify protein post Ni-NTA affinity chromatography, indicating how this system can be adapted to other purification types. Due to its setup with multiple columns, individual customization of parameters can be made in the same purification, unachievable by the current plate-based methods. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
