SOLICITATION NOTICE
H -- Laboratory Testing Services
- Notice Date
- 3/14/2003
- Notice Type
- Solicitation Notice
- Contracting Office
- Department of Health and Human Services, National Institutes of Health, National Institute of Allergy & Infectious Diseases/AMOB, 10401 Fernwood Drive MSC 4811 Room 2NE70-A, Bethesda, MD, 20817
- ZIP Code
- 20817
- Solicitation Number
- NOI3038
- Archive Date
- 4/12/2003
- Point of Contact
- Nerissa Smallwood, Purchasing Agent, Phone 301-402-6548, Fax 301-480-3695, - Sylvia Robinson, Purchasing Agent, Phone 301-402-2284, Fax 301-480-3695,
- E-Mail Address
-
nsmallwood@niaid.nih.gov, srobinson@niaid.nih.gov
- Small Business Set-Aside
- Total Small Business
- Description
- 17. This is a combined synopsis/notice of intent to procure commercial items prepared in accordance with the format in Subpart 12.6, as supplemented with additional information included in this notice. This announcement constitutes the only solicitation; quotes are not being requested for solicitation number NOI3038. The incorporated provisions and clauses are those in effect through Federal Acquisition Circular 01-12. This acquisition will be processed under Simplified Acquisition Procedures (SAP) and is a small business set-aside (NAICS-541380). The Government intends to procure testing services from BaroFold Inc the only known distributor that holds the exclusive patent rights to ?High Pressure Refolding of Protein Aggregates and Inclusion Bodies;? Patent No: US 6,489,450 B2. The contractor is to determine the conditions under which two candidate proteins (Pfs48 and Pvs48) can be refolded. The proteins would be characterized by a variety of methods to determine the extent of refolding. I.) Preliminary Screen of Refolding Conditions: For each of the proteins, baseline experiments to establish the effect(s) of pressure, ionic strength, pH and temperature on yields of soluble proteins. These studies will use variations around ?baseline? conditions as follows: Temperature: 25C; pressure: 1500 bar; pH 8.0 redox conditions: equimolar reduced and oxifized glutahtione, at a total concentration of 1 mM: ionic strength: 10 mM Tris, 40nM NaCl: protein concentration: 1 mg/ml. Four levels in addition to the ?baseline? will be used for each of the variables. For example, temperatures of 1, 10, 25, 40 and 50 C will be used, in solutions where all other variables are held at ?base case? values. Likewise, levels for pressure will be 1000, 1250, 1500, 2000, 2500 bar. Four levels of pH and ionic strength will also be chosen. In addition, selected consolvent conditions will be tested (e.g. addition of low levels of urea, guanidine HCL, or arginine). After contrifugation to remove insoluble fractions, soluble protein will be quantified by size exclusion chromatography, total protein assays (Bradford), and by SDS-PAGE using reducing and non-reducing conditions. For the conbination of refolding conditions that yield high concentrations of soluble protein, high-pressure spectorscopies (high pressure deriviative UV spectroscopy, high pressure fluorescence spectroscopy, high pressure light scattering) will be used to characterize structural changes in the proteins as they are processed from aggregated to soluble species. FTIR spectroscopy will be used to characterize protein secondary structure in the starting inclusion bodies, and compared with secondary structure measured by FTIR of the soluble fractions after high-pressure processing. Structure of the soluble protein will also be characterized by derivative UV spectroscopy, fluorescence, and far- and near-UV circular dichroism. In addition, correct formation of epitopcs in the resulting soluble fraction will be quantified using monoclonal antibody assays (Westerm Blotting and/or Elisa) in collaboration with NIH/NIAID researchers. The proteins of interest contain disulfide bonds. To increase the likelihood that the bonding pattern will result in correctly folded protein, the contractor will further refine these solution conditions by varying redox conditions. Ratios of reduced to oxidized glutathione ranging from 1:10 to 10:1 will be tested, at levels of disulfide scrambling agents of 0.5, 1, and 5mM. Similarly, other selected redox couples will also be tested. After pressurization and depressurization, levels of soluble protein will be determined. In addition, residula free cysteine in the soluble protein fraction will be quantified. Two or three samples from selected conditions will be analyzed by tryptic digests and mass spectrometry to ascertain patterns of disulfide bonds within the pressure-processed proteins depending on data results. For conditions of refolding/redox chosen, the contractor will measure the refolding kinetics and yields as a function of time and protein concentration. Mathematical engineering models of the refolding reaction kinetics will be developed to predict rates of refolding and applied in order to optimize production of native protein. These models will be compatible with the data generated for the specific protein. II.) Deliverables: A final report will be provided, within two weeks of completion of all lab procedures and data analysis, detailing the results of the experiments, and gives a description of attainable yields for high-pressure assisted refolding of each of the proteins. Recommendations for refolding conditions for each protein will be provided. Interim monthly updates will be provided with the monthly invoice for the work completed during that month. The government will provide to the contractor protein (Pfs48 and Pvs48). Written results will be reported to the government within four months from the delivery date of the protein. Government-designated personnel will address any problems noted directly to the contractor. Testing is to start mid April 2003 and continue through mid August 2003. The procedures that will be used in awarding this contract will be by written quotations or solicitations. The anticipated award date is March 28, 2003. This is a noncompetitive contract action set-aside for a small business. BaroFold, Inc. is exclusively licensed for this service. The following FAR provisions and clauses apply to this acquisition: FAR 52.212-1 Instructions to Offerors Commercial Items; FAR 52.212.3 Offeror Representations and Certifications Commercial Items; FAR 52.212-4 Contractor Terms and Conditions Commercial Items; FAR 52.212-5 Contract Terms and Conditions Required to Implement Statutes or Executive Orders Commercial Items. A completed copy of provisions at FAR 52.212-3 Offerors Representations and Certifications Commercial Items must be included. Copies of the aforementioned clauses are available upon request by telephone to Ms. Nerissa Smallwood at (301) 496-2607. Offerors must include with their offer a completed copy of the provisions at FAR 52.212-3, Offerors Representations and Certifications ? Commercial Items. Offers must be submitted no later than 5:00 PM Eastern Standard Time (EST), March 28, 2003 to Nerissa Smallwood. For delivery through the Postal Service, the address is NIH/NIAID/AMOB, 10401 Fernwood Drive, Suite 2NE70, MSC/4812, Bethesda, Maryland 20817-4812. Electronic submission will not be accepted. Requests for information concerning this requirement are to be addressed to Ms. Nerissa Smallwood at (301) 496-2607. Collect calls will not be accepted. All responsible sources may submit an offer that will be considered by this agency.
- Place of Performance
- Address: 5640 Fishers Lane, Rockville, Maryland
- Zip Code: 20852
- Country: USA
- Zip Code: 20852
- Record
- SN00279666-W 20030316/030315001116 (fbodaily.com)
- Source
-
FedBizOpps.gov Link to This Notice
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