REDACTED JOFOC 2177674.pdf

Justification for Other than Full and Open Competition at or Below the Simplified Acquisition Threshold

Project Title: CITE-seq analysis of Central spinal fluid cells in Multiple Sclerosis

I. Identification of the agency and contracting activity

DHHS, NIH, NIAID,

Requestors name:

Requestors phone number:

Requestors email address:

Address of the requesting lab/office: 10 Center Dr, Bldg 10, Bethesda, MD 20892

II. Description of the project, and supplies or services required to meet the agency’s needs (including the estimated value) A. Type of action. Check the appropriate box:

☒ New requirement ☐ Follow-on

• Name of the previous contractor(s):

Sequence number(s), if available:

• Was the initial and any subsequent awards competed? ☐ Yes ☐ No

B. AMBIS Sequence number for this requirement:

C. Total estimated dollar value of the requirement (including options, if applicable):

D. Period of performance, inclusive of options:

E. Project Background

The immune system in the blood does not reliably represent inflammation in the central nervous system (CNS) tissue.

To understand the phenotype of immune cells that play pathogenic role in CNS neuroimmunological diseases, one must study immune cells in the cerebrospinal fluid (CSF). However, even in patients with CNS neuroimmunological diseases such as multiple sclerosis (MS), the concentration of immune cells in the CSF is low: e.g., from 20cc of CSF (which represents close to maximum volume that can be safely collected in all adult humans) we can isolate approximately 20,000 immune cells. This is too few to perform mechanistic experiments.

However, novel technologies, such as scCITE-Seq (single-cell Cellular Indexing of Transcriptomes and Epitopes by sequencing) technique, which is developed and commercialized by 10x Genomics Inc., provide simultanous analysis of large number of surface epitopes together with RNA sequencing (Stoeckius et al, Nature Methods 2017). This is currently the only available methodology that can provide such deep characterizations of such rare immune cells we can safely collect in living humans with neuroimmunological diseases and controls.

Therefore, the goal of this project is to analyzing large number of patients with neuroimmunological diseases and controls collected under NDS natural history protocol 09-I-0032 with CSF scCITEseq to determine which cellular/phenotypic abnormalities are associated with what disease condition. We expect that reproducible/validated CSF cellular/transcriptomic abnormalities we can identify from this project will elucidate novel disease mechanisms that can be translated into new, more effective treatments.

There is currently only one available source of antibodies compatible with 10xGenomics CITEseq and these are BioLegend TotalSeq antibodies.

Because we plan using this technology on large cohort of patients, collected over several years, this will be a recurring order.

F. Project Description The main object of this research is to develop and optimize scCITEseq methodology applicable for longitudinal prospective analysis of cell surface markers and scRNAseq of CSF cells in MS and controls. We expect to analyze, in the blinded fashion, up to 100 CSF samples/year, up to 20,000 CSF cells/sample. We also expect that we will need total 200-300 samples to perform training cohort analyses and to validate major findings in the independent validation cohort.

The new produst from 10x Genomics make the CITE-seq techniq possible to analyze for the fixed cells. By using this new product: Chromium Single Cell Fixed RNA Profiling, we can use six samples per channel and reduce the running cost of CITE-seq. We plan to order the antibody for surface protein compatible with CITE-seq based on Chromium Single Cell Fixed RNA Profiling.

III. Identification of the statutory authority permitting other than full and open competition (SELECT ONE)

☒ 41 USC 253(c) (1) a n d F A R P a r t 13. 1 06 - 1 : Only one responsible source

IV. Demonstration that the proposed contractor(s) unique qualifications or the nature of the acquisition requires use of the authority cited.

A. Name and address of the proposed contractor(s) Biolegend 8999 BioLegend Way, San Diego, CA 92121, US Phone: (877)-246-5343 Web Site: https://www.biolegend.com

B. Describe why only the requested supply/service will meet the government’s needs There are only two sources of antibody products for cell surface protein which are available for CITE-seq analysis supplied by 10x Genomics: Biolegend Totalseq A and B. Among them, only Totalseq B is supported for CITE-seq analysis compatible 10x genomics on the fixed cells (Chromium Single Cell Fixed RNA Profiling). The technical protocol of Biolegend Totalseq B use in CITE-seq analysis was well established and showen in several reports (Kleino et al, Biotech J 2022 and Chen et al, Nat Commun 2021).

C. Describe why only the proposed contractor is capable of providing the requested supply/service The antibody for CITE-seq analysis must be barcorded with specific oligomer sequences to connect the surface protein expression with RNA information. Hence, Biolegend is only supplier for the antibody compatible with CITE-seq analysis.

V. Determination by the Contracting Officer that the anticipated cost/price to the Government will be fair and reasonable.

The price is determined to be fair and reasonable based on (provide a narrative describing how the determination was made and the rationale for the determination): See File, Price Reasonable Determination

VI. Certification

This is to certify that the justification for the proposed acquisition has been reviewed and that to the best of my knowledge and belief the information and/or data provided to support my request (for the requestor) and approval (for the CO) of is accurate http://www.biolegend.com/ and complete.