GE-HEALTHCARE ÄKTAdesign Purification Method handbook

Protein Purification Strategy

This handbook is intended to help you make full use of your ÄKTAexplorer™ or ÄKTApurifier™ chromatography system.The handbook is a collection of useful step-by-step protocols to aid your everyday purification work. Detailed instructions and recommendations are presented in a straightforward format, which should be easy to follow without needing a high level of expertise in programming or in chromatography.

This handbook is intended to help you make full use of your ÄKTAexplorer™or ÄKTApurifier™ chromatography system. Rather than being a text book in separation science, the handbook is instead a collection of useful step-by-step protocols to aid your everyday purification work. Detailed instructions and recommendations are presented in a straightforward format , which should be easy to follow without needing a high level of expertise in programming or in chromatography.Protocols specially adapted for the chromatography software UNICORN™ help you to plan, optimize and run your purification experiments.All purification protocols are based on pre-constructed templates available in UNICORN. In combination with the UNICORN Column list , these templates make method creation very simple.The handbook covers purification of proteins, peptides and oligonucleotides. However, before developing a suitable purification protocol, a development platform must be established, including:

• Required quality and quantity of the purified target molecule.

• Available information on the physical properties of the target molecule.

• Stability range for the starting material as well as the target molecule.

• Properties of the starting material.

The development of a suitable purification protocol starts with running Standard Purification Protocols. These protocols take advantage of the high overall resolution obtained by combining techniques with independent selectivities. This will provide sufficient purity in a large number of cases. When satisfactory results are not obtained with the Standard Purification Protocols, different optimizing measures are suggested.

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