SOLICITATION NOTICE
A -- Protein Extraction from Cell Membranes
- Notice Date
- 7/1/2002
- Notice Type
- Solicitation Notice
- Contracting Office
- Department of Energy, Pacific Northwest National Laboratory - Battelle (DOE Contractor), PNNL Licensing, PO Box 999, MSIN K9-89, Richland, WA, 99352
- ZIP Code
- 99352
- Solicitation Number
- Reference-Number-13569-E
- Response Due
- 9/1/2002
- Archive Date
- 9/16/2002
- Point of Contact
- Connie Mitzel-Faulk, Licensing Staff, Phone (509) 375-6401, Fax (509) 375-6731,
- E-Mail Address
-
technology@pnl.gov
- Description
- Pacific Northwest National Laboratory (PNNL), operated by Battelle Memorial Institute under contract to the U.S. Department of Energy, solicits interest from companies interested in obtaining license rights to commercialize, manufacture and market the following technology. License rights may be issued on an exclusive or nonexclusive basis and may include specific fields of use. PNNL may also be available to licensee(s) to assist in further research and development of the technology under a sponsored research agreement or CRADA program. The Technology: Membranes are critical components of cellular structure and function involving the partitioning of organelles, protecting the integrity of the genome, and providing defense from foreign molecules and external conditions that may damage or destroy the cell. Proteins that are inserted into the membrane are highly important biological and pharmacological targets. Despite their biological importance and natural abundance, the analysis of hydrophobic membrane proteins has been difficult. Techniques using two-dimensional polyacrylamide gel electrophoresis based mass spectrometry (2-D-PAGE-MS) often result in under-representation of these proteins. This is probably due to the inability of surfactants to effectively solubilize these water insoluble proteins in the aqueous medium used for isoelectric focusing. Moreover, these proteins tend to precipitate at their isoelectric point. Attempts to extract membrane proteins using organic solvent precipitation prior to 2-D electrophoresis have not shown significant improvement compared to surfactant-based protocols. An alternative technique to 2-D-PAGE-MS analysis is in-solution solubilization of the membrane proteins using surfactants followed by enzymatic or chemical fragmentation of membrane proteins, then separation and analysis of the complex mixture of peptides using liquid chromatography (LC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS/MS). However, the presence of surfactants can suppress analyte ionization and interfere with chromatographic separations, requiring additional manipulations to remove these contaminants and potentially lead to significant sample loses, particularly for low abundance proteins. Thus, the main obstacle for a large-scale mass spectrometric analysis of hydrophobic integral membrane proteins is the inability of current methodologies to achieve the dissolution of these hydrophobic proteins from the phospholipid bilayer while maintaining their solubility throughout the whole isolation and separation process and simultaneously avoiding the reagents which may interfere with ESI. To address this need, scientists at PNNL have developed a surfactant-free sample preparation method designed for large-scale LC-ESI-MS/MS analysis of integral membrane proteins. With this method, extraction of hydrophobic integral membrane proteins was achieved using high pH fractionation, thermal denaturation and organic solvent assisted solubilization. Additionally, avoidance of chaotropic denaturants and surfactants enhanced the quality of ESI-MS and further facilitated a large-scale LC-MS/MS analysis of the membrane subproteome. The results showed that hydrophobic membrane spanning peptides are readily detected using the developed method in conjunction with single-dimension high-pressure reversed-phase capillary LC coupled with ESI-MS/MS. Any company interested in licensing this technology must respond with a letter of interest (may be submitted by e-mail) no later than 30 days from the publication date of this Notice summarizing the company?s business and technical expertise and motivation for pursuing this opportunity. Companies deemed appropriate will be provided with further information on the technology. Such information may require an executed Nondisclosure Agreement. Respondents wishing to enter into negotiations for a commercial license will be required to submit a business plan for the commercialization of the technology prior to licensee(s) selection and negotiations. Please send letters of interest to the attention of the POC identified within this Notice.
- Record
- SN00103274-W 20020703/020701213351 (fbodaily.com)
- Source
-
FedBizOpps.gov Link to This Notice
(may not be valid after Archive Date)
| FSG Index | This Issue's Index | Today's FBO Daily Index Page |