I was stunned last week when I talking with an acquaintance of mine who is a Contract Specialist at SAIC-Frederick. I was stunned because I didn’t know he was in the Science business (technically, he’s more like a purchasing agent) and because he was complaining that he gets so little responses from local suppliers RFI and RFQ that many times these “Purchase Orders” just expire. I told him that I’d post some of these current opportunities, because I know many FredCoBio companies would love to have the chance to get a contract through SAIC.
Here’s the Beef. A full palette of delicious, juicy contracts just waiting to be bid upon. Viewable on FedBizOps:
S09-112 – Cell Line Development Services If no one jumps on this one, I’ll scream!!
Response Date: May 1, 2009 by 4:00PM Eastern
Archive date: May 30, 2009
Classification Code: A – Research and Development NAICS: 541711
Set Aside: N/A
Contracting office address: SAIC-Frederick, Inc.
National Cancer Institute at Frederick
92 Thomas Johnson Drive, Suite 250
Frederick, MD 21702-1201
A contract manufacturing organization or supplier with the ability to economically develop a mammalian cell line expressing a therapeutic protein including but not limited to chimeric or humanized monoclonal antibodies, antibody fragment fusion proteins, or recombinant cytokines in an established cell line, such as CHO suspension cells, HEK 293 cells, or NS0 cells with stable expression level at 10-50pg/cell/day using a batch, fed-batch, or perfusion process.
Potential respondents are to provide detailed description of technical approaches and experience, as well as estimated costs in performing the activities detailed below:
1) SAIC-F would provide a gene sequence of an antibody or antibody fragment fusion with a protein, or a recombinant protein with construct size <10-12KB.
2) Respondents will provide description of sequence analysis and codons optimization for expression in mammalian cells using either direct synthesis or codon optimization techniques.
Product Expression Regulation
1) Respondents will provide a description of promoter; e.g., CMV or equivalent strong mammalian cell promoters.
2) Respondents will use a leading peptide to express the protein as secreted form.
3) Respondents will use a terminator or terminators to enhance target gene transcription.
4) Respondents will provide details on removal of introns to improve transcriptional yield where acceptable.
1) Respondents will describe a selection system (GS or dhfr-) to be used in the expression system. Alternatives to the use of a selection system may also be described by the respondent.
2) Respondents will describe an additional selection to facilitate enriching high producer may be described; however SAIC-F requires to use a system previously approved by FDA for clinical trials.
3) Full length plasmid sequencing is required. A high transgene copy number in the cell line is preferred but not required.
Cell Line Requirement
1) Respondents will describe the proposed host cell line, such as CHO suspension cells, HEK 293 cells, or NS0 cells.
2) Respondents will describe how the host cell lines are qualified for use.
3) Respondents will provide how sequencing of the promoter and transgene coding region in the selected high producer is performed. Messenger RNA sequence and information regarding the location of the transgene(s) in the host cell is preferred but not required.
1) Respondents will describe procedures for how the constructed plasmid DNA is transfected into a selected cell line; to include:
a) A stable transfection pool.
b) Single cell cloning methods.
2) If a selection system is used in expression, a reasonable transfected clone number (depend upon the system) is required; please describe. If there is no selection system is used to enrich high producer, more than 1000 single clones are required to select for expression screening.
3) Responder will describe procedures to evaluate cell growth and productivity based on respondent’s selection on medium and culture condition.
Cell Growth and Productivity Requirement
1) Respondents will describe how the following that would be delivered:
a) A serum-free or protein-free medium adaptation.
b) Cell growth to reach 5-10 a 106 cells/mL.
c) Cell doubling time in the range of 20-30hour.
d) Viability of culture greater than 90%.
e) More than 5 days for batch process or more than 9 days for fed-batch process.
f) Productivity of greater than 10pg/cell/day is required.
2) Respondents will describe methods used to characterize the expression candidate protein or previous experience working with clients to achieve characterization.
Key Requirements to be considered for the areas above include:
1) A traceable history of the host cell used in the study.
2) A clear history of the plasmid vector used in the study.
3) A cell culture medium that is GMP compliant.
4) Generation of a cell line expressing a product is well documented.
5) All the sequence information of the plasmid and cell line to be made available in standard electronic formats.
S09-135 Endotoxin Production
Purpose of RFI:
SAIC-F is to seeking to identify a qualified contract research organization that is able to economically produce a new batch of clinical grade bulk endotoxin that is compliant with current Good Manufacturing Practices to replace the > 30 year old bulk endotoxin currently in storage. PLEASE NOTE: An appropriate strain of E. coli would be required to be acquired by any potential Subcontractor should a Request for Proposal (RFP) and subsequent Subcontracting Agreement be offered to a selected offeror.
The goal included in this statement of work is to produce a new batch of clinical grade bulk endotoxin that is compliant with current Good Manufacturing Practices to replace the > 30 year old bulk endotoxin currently in storage. This project will be broken down into a Development Section, including MCB production, and purification process development. An engineering run of bulk endotoxin along with a pilot lyophilization study. The final stage includes cGMP Production; including GMP bulk endotoxin manufacture and GMP vialing and lyophilization plus a manufacturing report.
Anticipated Milestones and Deliverables:
Milestone 1 – Activity A: Production and characterization of a 200 vial Master Cell Bank.
The previous cGMP endotoxin was produced from the Escherichia coli (Braude strain) group O 113:H10:K negative strain that was isolated and characterized at the Bacteriology Division, Bureau of Laboratories, Center for Disease Control, Atlanta, Georgia (Ewing WH, Hucks MC, and Taylor MW (1952) J Bacteriology 63: 319-325). The BDP will contact Dr. Epstein to determine if this bacterial strain is available from the NIH Clinical Center, the FDA, or the CDC.
The potential Subcontractor will obtain the appropriate strain of E. coli and then manufacture and characterize the 200 vial MCB according to current standards. The potential Subcontractor will provide a list of tests to be performed. SAIC-F may provide the potential Subcontractor a source of the required strain if necessary.
Milestone 1 – Activity B: Purification Development.
R&D grade E. coli cultures will be produced using the previously recommended chemically defined media and conditions. These cultures will be produced in the potential Subcontractor’s laboratories using standard R&D procedures and it is expected that this process will require minimal development work.
The resulting cell pellets will be used for purification development following the previous purification process used > 30 years ago. The potential Subcontractor will provide a proposal for process modification (if any) to meet the required scale, cGMP compliance, and product quality. Such studies may include:
1. A 10 liter culture will be produced and subjected to phenol extraction to produce a single unpurified bulk that can be used for purification development studies.
2. Tests to replace a dialysis step with transmembrane flow filtration (TFF).
3. Determine whether the deoxyribonuclease digestion can be replaced with Benzonase digestion.
4. Testing chromatography as a replacement for the sodium acetate/ethanol concentration step.
5. Determine whether the final dialysis step can be replaced with TFF.
Milestone 1 – Activity C: R&D Assay Development.
The potential Subcontractor will identify any method development necessary to perform the required release testing for the bulk and final vialed product shown in Tables 1 and 2.
SPECIAL NOTE: SAIC-F, with the NCI, will contact the US FDA for review of the proposed methods of analysis and specifications prior to initiation of the cGMP phase of the program. The results of that future discussion will be shared with the potential Subcontractor.
Milestone 2: Engineering Run/Pilot Lyophilization
The potential Subcontractor’s laboratories will be used to perform a cGMP pilot run. A full scale production and purification will be performed to test the process and produce bulk drug substance. This drug substance will be used as an internal reference standard for the following studies:
1. QC assay development.
2. QC testing for comparability to previous endotoxin CC-RE-Lot3.
3. Drug Substance stability studies.
The potential Subcontractor may perform any or all of these studies. The potential Subcontractor will also perform pilot formulation and lyophilization studies based on requirements to produce final vialed product listed in Table 3.
Milestones 3 &4: Production of cGMP Endotoxin
The potential Subcontractor will perform a full scale production and purification to produce cGMP grade endotoxin including the following activities:
1. Controlled fermentation will be conducted in a batch mode.
2. Purification will be performed to produce bulk drug substance.
3. QC/QA release of the drug substance.
4. Formulation, vialing, and labeling of endotoxin drug product (FVP).
5. QC/QA release of the FVP.
6. Stability program for FVP and, if required, bulk biological substance
7. Preparation of a Manufacturing Report (Appendix 1) to be included in
submissions to the US FDA with authorization to cross-reference any required
potential Subcontractor submissions to enable full review of the manufacturing
and testing documents by the US FDA for IND studies.
Table 1: Methods of Analysis and Specification for Bulk Biological Substance
Method Product Specification
Kinetic – QCL Potency Assay (EU/mL) (3) NLT 50,000 EU/mL
Gel Clot Potency Assay (EU/mL) (3) NLT 50,000 EU/mL
Sterility(1) Negative per 21CFR610.12
Residual host cell protein For Information Only
Residual host DNA For Information Only
Residual phenol(2) For Information Only
Mass spectrometry; method to be proposed by Contractor For Information Only
Polysaccharide characterization; method to be proposed by Contractor For Information Only
(1) Bacteriostasis & fungistatsis analysis to be performed on representative sample
(2) Additional residuals analysis may be required based on the potential process-related contaminants.
(3) Stability indicating assay
Table 2: Methods of Analysis and Specification for Final Vialed Product
Method Product Specification
Kinetic – QCL Potency Assay (EU/vial) 5,000 – 15,000 EU
Gel Clot Potency Assay (EU/vial) 5,000 – 15,000 EU
Sterility Negative per 21CFR610.12 & cUSP
Residual moisture <=3.0%
Note: All methods are stability indicating assays
Table 3: Required Deliverables for Milestone 3 & Milestone 4
CGMP Production Stage Minimum Quantity Configuration
Final Bulk Biological Substance 1.25 x 108 EU from 62.5 liters Final buffer is 1% lactose, 0.1% PEG-6000 to be bulk filled into sterile glass 10L bottles (8 L per bottle) and stored at 2-8oC
Final Vialed Product, Lyophilized 10,000 vials, net stability and release sampling; 2,000 EU/mL when reconstituted with 5 mL Water-For-Injection (WFI) 10,000 EU per vial in Type II borosilicate glass vials with 20mm stoppers and 20 mm West Aluminum seals. Sterile, single use vials stored at 2-8oC
S09-117 – DNA Plasmid Development Services
A contract manufacturing organization or supplier with the ability to develop a process, and perform manufacturing for DNA plasmids for therapeutics and vaccines, using an established cell line, such as E. coli DH 5 alpha with productivity of not less than 800mg/liter of purified plasmid DNA in fermentation broth. It is also required to demonstrate that high quality of plasmid product including but not limited greater than 90% of supercoiled DNA can be purified from the broth.
Potential Offerors are to provide detailed description of technical approaches and experience, as well as estimated costs in performing the activities detailed below:
Standard Plasmid Construct – Assume a gene insert would be provided to the Offeror for the purposes of responding to this Request for Information/Sources-Sought Notice:
1) Plasmid contains several basic components including but not limited to promoter(s) to allow gene expression in a mammalian cells and/or tissues; with bacterial replication capability in either high or low copy number, and with no beta-lactmase resistance marker.
2) Plasmid with size of less than 12Kb.
3) Plasmid carries at least one protein coding sequence for expression in mammalian cells and/or tissues.
Production System and Productivity
1) E. coli DH 5 alpha or equivalent FDA accepted E. coli strain that can be employed as host cell for production of a plasmid.
2) The E. coli cells used for production meet FDA requirements for GMP manufacturing. The Offeror is to provide standard listing of procedures to establish and characterize Master Cell Bank.
3) It is required to demonstrate that the productivity is 800mg/liter or higher of fermentation broth at 10 liter or larger fermentation scale. The Offeror is requested to provide experience with specific data.
4) It is required to demonstrate at small scale that the plasmid can be purified from the fermentation broth at reasonable purification yield. The Offeror is requested to provide experience with specific data.
Product Characterization and Quality Requirements
1) A full length plasmid DNA sequence is required; the Offeror is to provide methods used and identify any subcontract relationships, if applicable
2) It is required to demonstrate that the purified plasmid is acceptable in quality including but not limited to higher than 90% of supercoiled plasmid.
1) The Offeror is to provide facility and operational description for production of purified plasmid DNA under GLP and/or cGMP conditions. Examples of previous experience are requested.
2) It is requested that production be demonstrated for at least 5 g purified plasmid DNA per batch. The Offeror is requested to provide information on experience for large scale DNA plasmid production.