Inactive

The purpose of this purchase order is to acquire services to perform targeted affinity maturation of an anti-human BCMA nanobody via phage display library construction and selection. The objective is ... The purpose of this purchase order is to acquire services to perform targeted affinity maturation of an anti-human BCMA nanobody via phage display library construction and selection. The objective is to deliver five affinity-optimized nanobody sequences along with sufficient purified protein for in vitro and in vivo testing.

Inactive

The National Cancer Institute (NCI)’s Center for Cancer Research Office (CCR) Molecular Imaging Branch (MIB) plans to procure on a sole source basis services for targeted affinity maturation of an ant... The National Cancer Institute (NCI)’s Center for Cancer Research Office (CCR) Molecular Imaging Branch (MIB) plans to procure on a sole source basis services for targeted affinity maturation of an anti-human BCMA nanobody via phage display library construction and selection. This acquisition will be processed in accordance with simplified acquisition procedures as stated in FAR Part 13.106-1(b)(1) via request for written quotation. Only one (1) award will be made as a result of this solicitation. This will be awarded as a firm fixed price type contract. The Period of Performance is estimated to be a base period of six (6) months from the effective date stated in the award. 2.0 BACKGROUND The National Cancer Institute/Center for Cancer Research (NCI/CCR), Molecular Imaging Branch (MIB) is engaged in the development of radiolabeled nanobodies targeting clinically relevant tumor antigens for diagnostic and therapeutic applications. One of the current programs focuses on targeting B-cell maturation antigen (BCMA) in multiple myeloma. A parental anti-human BCMA nanobody was purchased and demonstrated promising preliminary binding in vitro and in vivo, but further affinity enhancement is required to support translational applications. The goal is to generate high affinity nanobody variants (based on the parental one), suitable for imaging and radiotherapy through affinity maturation using phage display. 3.0 OBJECTIVE The Contractor shall perform targeted affinity maturation of an anti-human BCMA nanobody via phage display library construction and selection. The objective is to deliver five affinity-optimized nanobody sequences along with sufficient purified protein for in vitro and in vivo testing. 4.0 SCOPE The Contractor shall perform nanobody affinity maturation using established phage display techniques. The project will be based on nanobody sequence provided by the Government and will encompass a series of integrated activities aimed at enhancing binding affinity to the BCMA target antigen. The Contractor will initiate the work by producing recombinant nanobody protein in a mammalian expression system to confirm expression and proper folding. The binding activity of the parental nanobody to BCMA will be evaluated using immunoassays such as ELISA, followed by quantitative affinity measurements using validated platforms (e.g., BLI or SPR). Subsequently, the parental nanobody will be cloned into a suitable phagemid vector for display and functional validation. Mutant libraries will be constructed through in silico-guided design, utilizing error-prone PCR and semi-rational mutagenesis to introduce diversity across relevant regions of the nanobody. Affinity selection will be conducted through multiple rounds of biopanning against soluble BCMA antigen to enrich for variants with improved binding characteristics. Candidate clones from enriched libraries will be screened via ELISA or equivalent binding assays, and selected high-affinity binders will be sequenced and analyzed. Top-performing nanobody variants will be recombinantly expressed in mammalian cells, purified, and subjected to standard quality control procedures. Binding affinities of the matured clones will be re-evaluated to confirm improvements over the parental nanobody. Up to five optimized nanobody clones will be delivered to the Government, including the corresponding amino acid and nucleotide sequences. These five clones will be provided to the MIB for downstream evaluation. At least one of the five delivered clones must demonstrate a minimum 10-fold improvement in binding affinity relative to the parental nanobody. The Government shall not exercise any supervision or control over the contract service providers performing the services herein. Such contract service providers shall be accountable solely to the Contractor who, in turn is responsible to the Government. The Contractor shall provide all personnel, labor, facilities, materials and equipment necessary to generate the synthesized compounds. All development and production will be performed off-site. 5.0 CONTRACT REQUIREMENTS / PERSONNEL QUALIFICATIONS The Contractor must have demonstrated expertise in nanobody engineering, phage display library construction, and affinity maturation, with a proven track record in delivering affinity-optimized nanobody reagents for therapeutic or diagnostic applications. The Contractor shall perform nanobody affinity maturation using phage display methods, based on sequences provided by the Government. The Contractor shall perform the following tasks: 1. Parent VHH Recombinant Production Based on the Government provided parental nanobody sequence, the Contractor shall produce 1 mg of purified VHH protein using mammalian cells (293 or CHO). 2. Parent VHH ELISA Validation Evaluate the binding activity of the recombinant VHH to the target BCMA. The Government-supplied VHH protein will serve as a control. 3. Parent VHH Binding Affinity Measurement Measure the binding affinity of the parent nanobody to the BCMA antigen using Octet BLI or Biacore SPR. The results shall be shared with the Government for decision-making prior to continuing. 4. Parent Phage VHH Construction and Binding Test Clone the parent VHH into a phagemid vector for phage display and validate for target binding. 5. Design and Construction of Mutant VHH Phage Display Libraries Based on in silico analysis of the parent VHH, construct multiple mutant libraries (VHmut-VLwt, VHwt-VLmut, VHmut-VLmut, and hotspot library) using error-prone PCR and semi-rational design approaches. 6. Biopanning Against Target Antigen Using Mutant Libraries Perform three to four rounds of in-solution panning using soluble BCMA antigen to enrich nanobody variants with increased affinity. 7. Phage ELISA and Validation Evaluate up to 300 phage clones from the enriched pool. Clones with superior binding will be selected, sequenced, and analyzed. 8. Recombinant Production of Selected Nanobodies Use mammalian systems to express the five top-ranked affinity-matured nanobody clones. Each will be produced at a scale of 2 mg, QC-tested by SDS-PAGE, and delivered to the Government. 9. Affinity Measurement of Matured Nanobodies Meausre the binding affinities of the five purified clones using Octet BLI or Biacore SPR to determine improved KD values. 10. Milestone 2 - Deliver up to 5 new clones (2mg from each), with at least 1 clone showing a 10-fold improvement in affinity. 11. Internalization Evaluation - Evaluate internalization performance on target Cells, and confirm the new nanobody sequences have better internalization than the parent nanobody used for the affinity maturation. 6.0 TYPE OF ORDER It is anticipated that this requirement will be a firm fixed price purchase order. 7.0 ANTICIPATED PERIOD OF PERFORMANCE The Period of Performance is estimated to be six (6) months base period from the effective date stated in the award. 8.0 PLACE OF PERFORMANCE Services shall be performed at the contractor’s facilities. 9.0 SOLE SOURCE JUSTIFICATION The tasks outlined in the Contract Requirements require that the contractor has demonstrated expertise in nanobody engineering, phage display library construction, and affinity maturation, with a proven track record in delivering affinity-optimized nanobody reagents for therapeutic or diagnostic applications. The goal is to generate high affinity nanobody variants (based on the parental one), suitable for imaging and radiotherapy through affinity maturation using phage display. Creative Biolabs originally supplied the BCMA-targeting nanobody used in NCI’s proof-of-concept studies. Their nanobody demonstrated the best binding performance in NCI’s in vitro and in vivo comparisons. Because they generated the parental nanobody and possess proprietary knowledge and tools used in its development, Creative Biolabs is uniquely qualified to enhance its affinity while preserving key binding characteristics. The goal of this acquisition is to generate a panel of improved BCMA-targeting nanobodies suitable for both imaging and therapeutic applications, in support of MIB’s broader mission to advance molecularly targeted agents toward clinical use. 10.0 SUBMISSION INSTRUCTIONS This notice is not a request for competitive quotation. However, if any interested party, especially small businesses, believes it can meet the above requirement, it may submit a capability statement, which shall be considered by the agency. The statement of capabilities and any other information furnished must be in writing and must contain material in sufficient detail to allow the NCI to determine if the party can perform the requirement. Information furnished must not exceed 5 pages (12-point font minimum) and should include an outline of previous or similar projects performed. Responses must be received in the contracting office by 3:00 PM ET, August 1, 2025. All responses and questions must be in writing and emailed to Michelle Nguyen, Contracting Officer, via electronic mail at michelle.nguyen2@nih.gov. Reference Notice Number 75N91025Q00049 on all correspondence. The Government’s determination to solicit quotations from the vendors to compete this proposed requirement, 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. In order to receive an award, Contractors must be registered and have valid, current Entity Record, including current Representations and Certifications, in the System for Award Management (SAM) through SAM.gov. 11.0 DISCLAIMERS AND IMPORTANT NOTES This notice does not obligate the Government to award a contract or otherwise pay for the information provided in response. The Government reserves the right to use the information provided by respondents for any purpose deemed necessary and legally appropriate. Any organization responding to this notice should ensure its response is complete and sufficiently detailed to allow the Government to determine the organization’s qualifications to perform the work. Respondents are advised the Government is under no obligation to acknowledge receipt of the information received or provide feedback to respondents with respect to any information submitted. Responses to this notice will not be considered adequate responses to a solicitation. Confidentiality: No proprietary, classified, confidential, or sensitive information should be included in your response. The Government