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

99 -- PRODUCE MONOCLONAL ANTIBODY-35, (MAB-35) PURIFIED BULK AND CONJUGATE MAB35-RICIN SOL RFQ-NCI-00145-NV DUE 071700 POC Debbie Moore, Purchasing Agent (301) 402-4509, Todd Cole, Contracting Officer (301) 402-4509 This is a combined synopsis/solicitation for commercial services, prepared in accordance with the format in FAR 12.6, as supplemented with additional information included in this notice. This announcement constitutes the only solicitation and a separate solicitation will not be issued. This solicitation, No. RFQ-NCI-00145-NV includes applicable provisions and clauses in effect through FAR FAC 97-17. The acquisition is being made in accordance with the test program for using simplified procedures for certain commercial items authorized in FAR 13.5. The solicitation is set-aside for small business. The Standard Industrial Classification Code is 2836, and the business size standard is 500 employees. BACKGROUND: The National Cancer Institute (NCI), on behalf of the National Institute for Neurological Disorders and Stroke, requires monoclonal antibody-35. This project involves the production and delivery of an immunotoxin for clinical trials for the treatment of a variety of muscle spasm disorders. In animal models, use of an immunotoxin specific for the muscle acetlycholine receptor is effective in reducing muscle strength (see Neurology Hott, JS, Dalakas, MC, Sung, C, Hallett, M and Youle RJ: A skeletal muscle-specific immunotoxin for the treatment of focal muscle spasm. Neurology 50: 485-491, 1998). REQUIREMENTS: Independently, and not as an agent of the Government, the Contractor shall perform the commercial services described below and provide all personnel, material, equipment, and facilities not otherwise specified and provided by the Government. Specifically, the Contractor shall, under Good Manufacturing Practices (cGMP), produce monoclonal antibody-35, (Mab-35) purified bulk and conjugate Mab-35 to Ricin supplied by NIH, purify the conjugate and make available container products for phase III clinical trials. Following reviews of validation and qualification documents for facility, systems, equipment, raw materials, and process used in this project, working banks of purified monoclonalantibody -35, and purified Mab-35 conjugate to Ricin shall be prepared and qualified and stored on-site at the Contractor's facilities. Mab-35 production and conjugation to Ricin shall be performed at an FDA-approved clinical manufacturing site. Two production lots of the Mab-35 / Ricin shall be produced under cGMP guidelines as the primary final bulk product and a backup bulk lot. If the backup lot is not needed for production of final container product, then the backup lot shall serve as a control lot for QC testing and archives and as a validation lot. Mab-35 shall be purified from mouse ascites by ammonium sulfate precipitation and DEAE Sepharose chromatography. Conjugation of Mab- 35 to Ricin shall be performed by adding DTT in phosphate-buffered saline to antibody, and incubating for 30 minutes. This antibody-DTT mixture will be applied to a G-25, gel filtration column equilibrated with PBS. Peak antibody fractions will be collected and pooled. Then, Ricin will be mixed with DMF containing the bi-functional cross-linking agent, MBS and incubated at room temperature for 30 minutes. The Ricin-MBS will be immediately reacted with fresh reduced antibody and incubated at 4 degrees overnight. The conjugate will then be separated from unreacted ricin by HPLC on a TSK 3000 SW column in sodium phosphate buffer (pH 7.2), or another size exclusion column. The peak fractions from several runs containing both unreacted antibody and ITX (immunotoxin) will be pooled and loaded onto an immobilized D- galactose affinity column at 4 degrees. After the column is flushed with PBS to remove unreacted antibody, lactose will be run over the column to elute the purified ITX. Each GMP lot shall contain at least 100mg. of purified Mab-35-ricin ITX. The final bulk products shall undergo analytical and safety testing by the Contractor's quality control group and the SNB of NINDS at the National Institutes of Health, prior to release for formulation and final container filling. Review and approval of batch records and test results shall be performed by the Contractor's Quality Assurance (QA) group. Aseptic formulation and filling of final container vials shall be performed at an FDA-approved clinical fill-finish operation. Upon, QA release, the final bulk product shall be formulated at 0.1 mg/ml in PBS (pH7.2), and Human Serum Albumin at 1mg/ml, and dispensed into 500 final container vials at 2 ml per vial. The final bulk and container products shall be stored at 4 degrees in secure refrigerators at the Contractor's location. The final container products shall be subjected to safety and analytical testing conducted by the Contractor's Quality Control group and the SNB of NINDS at the National Institutes of Health. Quality Assurance by the Contractor shall encompass reviewing of batch records, final container test results, and final manufacturing reports. Upon approval, the Contractor shall issue certificates of analysis and release the final container products. Final manufacturing reports shall be provided to the NINDS or designated entity as noted by the Project Officer. The final bulk and final container products shall be enrolled into a stability testing program for the Mab35-ricin conjugate ( ITX) . Upon written notification from the Project Officer, the Contractor shall ship at 4 degrees, the final container products to the SNB/NINDS repository in Bethesda, Maryland. The Contractor shall adhere to the following objectives and timelines in completing the above work: MONTH 1 -- 1.Review validation and qualifications of off-site manufacturing plant; 2. Implement technology transfer process of off- site manufacturing and fill-finish. Facilities; 3. Review SOPs and prepare batch records for GMP production of Mab35 & Mab35-ricin conjugate; 4. Produce 200 ml of Mab35 ascites as a working bank from NIH Swiss Nude mice. MONTH 2 -- Purify Mab35 ascites by ammonium sulfate precipitation and DEAE Sepharose Chromatography, in manufacturing cleanroom by following processing scheme below : a.mix ascites with an equal volume of cold saturatedammonium sulfate, and stir at 4 degrees for 20minutes; b.centrifuge ascites mix in Sorvall 34 rotor at 8000 rpm for 10min., remove supernate and save at 4 degrees C. (Fraction S); c. resuspend ascites pellet (Fraction P) in 20mM Tris (buffer A), pH 7.9; d. dialyze Fraction P against 2 changes of 2L of 20mM Tris buffer, pH 7.9 containing 40mM NaCl (Tris buffer D), at 25 C over a 4hr. period or against Tris Buffer A . Repeat dialysis with fresh 2L buffer overnight; e. Remove sample from dialysis bag & place in clean glass centrifuge tubes; f. Centrifuge in Sorvall 34 rotor at 8000 rpm for 10 min. to remove any aggregated protein. (save 25 ul for SDS gel); g. Apply fraction P over pre-poured & packed DEAE-sepharose column and elute with a sodium chloride gradient, (buffer B & buffer C) low & high salt. Collect fractions from column gradient in tubes of 2ml on fraction collector. (buffer B= 20mM Tris+20mM NaCl) & (buffer C=20mMTris + 400mM NaCl); h. Read fraction tubes on spectrophotometer at A280 and store tubes at 4 degrees until needed; 2. Analyze Mab35 fractions collected by Quality Control methods at the Contractors; and at SNB, NINDS of the National Institutes of Health using the following: a. Perform SDS-PAGE protein analysis on fraction A, (pre-dialyzed ascites) Fraction P, ( dialyzed Mab35 pre column), a MW standard and fractions of interest from column gradient samples; b. PAGE should be run using Tris-Glycine 1x buffer; c. Gel should be stained with Coomassie Blue Reagent and destained with 10% acetic acid; d. Using A280 results and gel samples as a guide, pool fractions with highly purified IgG and dialyze and/or concentrate; e. Dialyze against 2L PBS ph 7.4 and change PBS buffer 1x over a 2-4 hr. period / or overnight to remove sucrose; f. Remove sample from bag as carefully as possible and place in clean tube; g. Centrifuge in clean tubes in Sorvall rotor for 10min at 8000 rpm; h. Pour supernate into clean sterile tubes and calculate concentration of Mab35 antibody. MONTH 3 -- 1. Perform conjugation of Mab35 antibody to ricin by following methods: a. Add DTT in phosphate- buffered saline (PBS) to 10-50mg of antibody, and incubate for 30 min to partially reduce the antibody; b. Apply this antibody-DTT mix to a G-25, gel filtration column equilibrated with PBS, collect fractions and pool peak antibody fractions; c. Mix ricin, supplied by NIH, 20- 100mg in PBS with DMF containing the bi-functional cross-linking agent, MBS (m- maleimidobenzoyl-N-hydroxysuccinimidyl) ester; d. Incubate mixture at room temperature for 30 min. Ricin-MBS is then immediately reacted with freshly reduced antibody and incubated at 4 overnight; e. The conjugate is then separated from unreacted ricin by HPLC on a TSK 3000 SW column in 0.1M sodium phosphate buffer (pH 7.4), or another size exclusion separation method; f. The peak fractions from several runs containing both unreacted antibody and ITX (immunotoxin) shall be pooled and loaded onto an immobilized D-galactose affinity column. (Pierce) at 4 degreesCelsius; g. The column is then flushed with PBS to remove unreacted antibody, and 0.1M lactose is run over the column to elute the purified ITX. MONTH 4 -- 1. Review of QA batch records and testing and release of final bulk product; 2 . Formulate final bulk product in Sterile PBS and Human Serum Albumin at 1mg/ml as before in glass tubes in cleanroom; 3. Perform filling of produced Mab35 and Mab35-ricin conjugate into glass vials. Each borosilicate glass vial shall contain 2 ml aliquoted aseptically in class 100 Biosafety hood in class 100 filling cleanroom. Gray butyl stoppers lined with Teflon will be inserted aseptically into vials. Aluminum caps will be crimped automatically using filtered nitrogen gas; 4. Label vials with sponsor (provided by Project Officer ) approved information; 5. Transfer labeled vials on 4 degree ice packs from fill-finish facility to NINDS; 6. Complete archival storage of final container product at 4 degrees celsiuis in a secure refrigerator for stability testing; 7. Performquality control safety testing of final container product including: a. microbial sterility testing, b. Mycoplasma and spiroplasma testing, and c. Endotoxin testing; 8. Perform QC analytical testing of final container product including: a. Purity-SDS-PAGE analysis with scanning densitometry and EM, b. Identity -- Western blot analysis, c. Strength-BCA protein quantitation assay; 9. Perform QA auditing of batch records and final manufacturing report, release of final container product, and issuance of certificate of analysis; 10. Prepare and submit final manufacturing report by project manager to sponsor; 11. Perform stability testing of final bulk and container product per existing stability program; and 12. Finally, in addition to the above, documentation of the production procedure and the specifications of the final material, along with certificates of the GMP techniques used throughout the process and bottling, shall be provided in sufficient detail to allow for the FDA review and approval. Shippingand Delivery: Mab35 and Mab35-ricin conjugate final container products shall be packaged according to the requirements herein. The products shall be shipped at 4 degrees to a location within Bethesda, Maryland area. The exact location shall be documented within the anticipated award document. All products shall be shipped at the same time. Furthermore, the products shall be shipped F.O.B. destination or F.O.B. origin with transportation insurance provided by the vendor with inside delivery to the location specified within the anticipated order. It is planned that final delivery of the product and all documentation will be prior to March 1, 2001. PRICING: Quotations must include a complete fixed price which includes the product, all certifications, warranties, reports, other fees/costs, shipping, and insurance, if F.O.B. origin. EVALUATION CRITERIA: The technical portion of quotations will be reviewed by a panel of at least three NIH experts. The technical portion of quotations will receive paramount consideration in selecting a vendor. However, cost/price will also be a determining factor in the event that two or more offers are determined to be essentially equal following the evaluation of technical factors. Vendors submitting quotations shall also provide three references of organizations that have produced similar monclonal antibodies within the past three years. The references may be used in the event that two or more offerors are considered essentially equal after the technical and price evaluations. In any event, the Government reserves the right to make award to the vendor whose offer provides the greatest overall value to the Government. The major evaluation factors for this RFQ include technical and price factors. MANDATORY QUALIFICATION CRITERIA: The following is a mandatory qualification criterion that establishes conditions that must be met in order for your quote to be considered for award: vendor/quoters must demonstrate that they own or have legal access to an FDA-approved clinical manufacturing site to perform the requirement herein. TECHNICAL EVALUATION CRITERIA: The following technical evaluation criteria will be used by the Government when reviewing the technical portion of the quotation. The criteria below are listed in the order of relative importance with weights assigned for evaluation purposes: 1. Technical understanding of the requirements to produce two production lots of Mab-35/Ricin under cGMP guidelines relative to technical approach, techniques, and quality assurance and control to ensure the required purity. (45%); 2. Evidence of having produced similar monoclonal antibodies under cGMP guidelines. In addition, availability and suitability of personnel to perform such monoclonal antibody production relative to training and experience. (40%); and 3. Suitability of facilities and equipment for production of Mab-35/Ricin under cGMP guidelines (15%). Total weighting of above factors is equal to 100%. Quotes must be submitted on an SF-18 or SF-1449. Quotes must be accompanied by a completed "Offeror Representations and Certifications- Commercial Items -- with Duns Number Addendum," signed by an authorized representative of the vendor. Quoters must submit an original technical proposal/quotation plus descriptive literature, warranties, and/or other materials that demonstrate that their offer meets all of the foregoing requirements. The technical proposal's documentation shall also include curriculum vitae for the contractor staff that will be involved in the production; legal proof of ownership or access to a GMP facility for producing the above material; and a narrative of experience in producing similar products via similar commercial processes. In addition, four (4) copies of the technical proposal and all materials shall be submitted with the original. This solicitation incorporates the provisions at FAR 52.212-1, INSTRUCTION TO OFFERORS -- COMMERCIAL ITEMS -- DEVIATION FOR SIMPLIFIED ACQUISITIONS; FAR 52.212-3, OFFEROR REPRESENTATIONS AND CERTIFICATIONS -- COMMERCIAL ITEMS, along with the DUNS NUMBER ADDENDUM; and The resulting contract will incorporate the requirements contained in paragraphs (a)and (d) of the clause at FAR CLAUSE 52.212-4 CONTRACTS TERMS AND CONDITIONS COMMERCIAL ITEMS and FAR Clause 52.212-5, CONTRACT TERMS AND CONDITIONS REQUIRED TO IMPLEMENT STATUTES OR EXECUTIVE ORDERS- COMMERCIAL ITEMS-DEVIATION FOR SIMPLIFIED ACQUISITION. The contract will also incorporate the following FAR clauses cited in paragraphs (b) and (c) of the latter clause: FAR 52-222-26, EQUAL OPPORTUNITY; FAR 52-222-35, AFFIRMATIVE ACTION FOR DISABLED VETERANS AND VETERANS OF THE VIETNAM ERA; FAR 52-222-36, AFFIRMATIVE ACTION FOR WORKERS WITH DISABILITIES; FAR 52-222-37; EMPLOYMENT REPORTS ON DISABLED VETERANS AND VETERANS OF THE VIETNAM ERA; and FAR 52.232-34, PAYMENT BY ELECTRONIC FUNDS TRANSFER OTHER THAN CENTRAL CONTRACTOR REGISTRATION. Interested vendors may contact Debbie Moore, Purchasing Agent, at (301)- 402-4509 (Fax: (301)-402-4513) to receive full text copies of the representations and certifications or other cited provisions and clauses. Any questions pertaining to this solicitation and requirement shall be submitted by July 10, 2000. Complete quotations and all related materials must be received in this office no later than 3:00 pm EDT on July 17, 2000. Please cite the solicitation number, RFQ-NCI-00145-NV, on your quotations, including all copies of the technical portion of quotation. Posted 06/29/00 (W-SN470146). (0181)

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