National Cancer Institute, Research Contracts Branch, PSAS, 6120 Executive Blvd, EPS/Room 638, Bethesda, MD 20892-7227

66 -- OCTOPUS UNIVERSAL MODULAR SOL RFQ80233-NS DUE 092198 POC Patricia Haun (301) 402-4509 The Laboratory of Cell Biology (LCB), Division of Basic Sciences (DBS), National Cancer Institute(NCI) plans to procure an Octopus-universal modular for the basic process of continuous free flow electrophoresis from GmbH Weber, Klausnerring 17, D-85551 Kirchheim-Heimstetten,Germany. The comparison of two dimensional-gel electorphoresis (2-DE) separations from extracts of primary tumors have revealed several proteins which are up or down regulated in different types of cancerous cells. There is an urgent need for unique tumor markers which can be utilized for correct diagnosis and treatment. Two dimensiona-gel electrophoresis offers high resolution of proteins present in cell extracts. In addition, when applied to the analysis of primary tumors, two dimension gel eletrophoresis provides unique spots that can be clearly detected. The Octopus-universal modular system, which is produced by GmbH Weber, is a preparative system which NCI can use for the purification of organelles and various proteins from crude mixtures prior to further separation by a second dimension on 2-D gel electrophoresis (2-DE). NCI requires high performance separations in the system with a resolution in the range of 0.02-0.04 pI. The problem of coelectrophoresing protein spots in 2-DE is well recognized and the approach is technically limited and capable of resolving and detecting only a few thousand of the most abundant ("house-keeping") proteins. To enrich for low-abundance proteins, it is necessary to analyze sub-cellular components (e.g., membrane proteins, lysosomes, and other organelles) which can be purified only by the Octopus system. The approach for identifying and characterizing proteins utilizes modern mass spectrometric techniques such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) mass spectrometry (MS)in combination with genome database searching strategies . By combining the Octopus system and 2-DE with ESI-tandem mass spectrometry (ESI-MS/MS) and, more recently, by utilizing nanoelectrospray ion source MS/MS, it is now possible to analyze complex mixtures of peptides at low femtomole levels to obtain sequence information. Using this approach, proteins present in simple mixtures can be identified over a 30-fold difference in molar quantity. However, the yield and purity of the fractions is crucial to a succesfully protein identification. Protocols are now established on the Octopus system using flat gradients made by a new type of media with a span of 1 or 2 pH units and a resolution in the range of 0.02 pI, that will allow purification and high yield of low amounts of protein samples from extracts of primary tumors. Requirements: The system shall include a basic instrument for continuous free flow electrophoresis, accessories for an IZE process, a cooler for the PZE process, a heat exchanger for temperature control, a high voltage power supply, a peristaltic pump, and maxi prep fractionation cabinets. In addition, the system must be capable of separating organelles and proteins to a degree of 90% purity acceptable for the aforementioned work. Moreover, using flat gradients, made of ampholytes with a span of 1 or 2pH units, the resolution shall be capable of being increased up to .02 pI. Since the Octopus meets the purity and the resolution (.02 pI) required for NCI's studies, GmbH Weber is only source known to NCI that can meet all of the above requirements. If any interested party believes it can perform the above work and can demonstrate, it may submit a statement of capabilities. The statement of capabilities and any other information furnished must be in writing and must contain material in sufficient detail to allow NCI to determine if the party can perform this requirement. Capability statements must be received in the contracting office by 3:00 p.m. EST, on September 21, 1998. If you have any questions, please contact Patricia Haun, Purchasing Agent on (301) 402-4509. A determination by the Government not to compete this proposed requirement based upon responses to this notice is solely within the discretion of the Government. Information received will be considered solely for the purpose of determining whether to conduct a competitive procurement. Posted 09/03/98 (W-SN245653). (0246)

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