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Optimum Protein Solubility Screen

IB-2005

APPLICATIONS OF TECHNOLOGY:

  • Protein preparation for x-ray crystallography and NMR
  • Protein purification kits and reagents
  • Protein functional analysis

ADVANTAGES:

  • Rapidly identifies optimal conditions for protein solubility
  • Works with very small amounts of protein (240 mg)
  • Readily adaptable for high throughput processing; easily scalable to larger volumes

ABSTRACT:

Sung-Hou Kim and colleagues at Berkeley Lab have developed a novel method that increases protein solubility. The method improves the success rate of both solubility and crystallization, critical steps in x-ray crystallography and Nuclear Magnetic Resonance (NMR).

Protein solubility is often the bottleneck phase of the purification process. Use of an optimum solubilization buffer is of key importance, but at present, buffer selection largely relies on educated guesswork.

The Berkeley Lab method enables rapid, optimum buffer selection by simultaneously screening very small amounts (240 mg) of protein for solubility in twenty-four buffers. Solubility is then assessed using a light microscope and dynamic light scattering (DLS). If the solubility is not yet satisfactory, a variety of additives can be further screened by DLS for improved solubility.

This method has been used by Berkeley Lab researchers to concentrate and crystallize previously problematic proteins.

STATUS:

FOR MORE INFORMATION:

Jancarik, J., Pufan, R., Hong, C., Kim, S., Kim, R. "Optimum Solubility (OS) Screening: An Efficient Method to Optimize Buffer Conditions for Homogeneity and Crystallization of Proteins," Acta. Cryst. 2004, D60, 1670-1673.

REFERENCE NUMBER: IB-2005

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