The Tetramer Facility provides 0.2 mg of MHC /peptide tetrameric reagent, which is enough for 200 assays.
Tetramers and/or their derivatives are provided for research purposes only. Their use for direct clinical applications or commercial purposes is prohibited.
Requests are made via the online Request Form which is available on this site. The NIH Tetramer Resource Committee (TRC) meets every two weeks to review, approve, and prioritize applications (see Request Prioritization). Requestors are informed of approval or disapproval of their request by E-mail. Receipt of final tetramers can take 2-3 months from time of initial request, depending on the TRC approval date and the workload of the NIH Tetramer Facility.
All requests received by 12pm ET on the day prior to the NIH TRC meeting will be reviewed at the next day's meeting. Incomplete requests or requests received after 12pm will be reviewed in approximately two weeks of date of request. The NIH TRC meets on the following dates in 2008: Jan. 4, Jan. 18, Feb. 1, Feb. 15, Feb. 29, March 14, March 28, April 11, April 25, May 5, May 30, June 13, June 27, July 11, July 25, August 8, August 22, Sept. 5, Sept. 19, Oct. 3, Oct. 17, Nov. 7, Nov. 21, Dec. 5, Dec. 19, and Jan 9, 2009.
Investigators may request up to 10 tetramers per 12 month period from the initial date of approval by the TRC. Please note that requests beyond 5 tetramers in one twelve month period will be approved pending availability of resources at the Tetramer Facility. Exceptions to this policy may be considered by the TRC under special circumstances.
The Facility provides MHC class I and CD1 tetramers labeled with streptavidin-allophycocyanin; streptavidin-phycoerythrin; streptavidin-Pacific Blue; streptavidin- Fluorescein; streptavidin-Alexa 488; streptavidin-Alexa 647; or streptavidin-Alexa 680. If other labeling reagents are desired, they must be provided by the requestor. Some lot to lot variation in streptavidin-phycoerythrin has been observed, but should not seriously affect tetramer sensitivity or background if the suggestions outlined in the provided protocol are followed. Pre-made and custom class II tetramers can be labeled with streptavidin-allophycocyanin or streptavidin-phycoerythrin.
The Facility can provide MHC class I tetramers with human or murine beta-2 microglobulin, upon request. The beta-2 microglobulin for CD1 tetramers is dependent upon the CD1 species requested, since the beta-2 and CD1 genes are expressed from the same plasmid.
If the requested MHC allele is not currently at the NIH Tetramer Facility or if there are other requirements, such as a specific beta-2 microglobulin needed to reconstitute Class I of an unusual species, the investigator must supply a clone. A current list of available MHC alleles is available on this site. The Facility will perform the subcloning into the appropriate vector for protein expression. All plasmids containing MHC alleles will be retained at the NIH Tetramer Core Facility and will not be distributed. These alleles will become property of the NIH Tetramer Core Facility and will be used for future production and distribution of tetramer reagents.
Quality control of tetramer reagents produced at the facility includes the following:
ELISA using anti-beta 2 microglobulin antibodies specific for properly folded MHC I.
FPLC chromatogram of the protein indicating its size to demonstrate proper folding and multimerization of the tetramer reagent.
Upon request approval, investigators will need to:
Send 10 (ten) milligrams of HPLC purified peptide for custom MHC class I tetramer synthesis or 0.5 milligrams for custom MHC class II tetramer synthesis, with its chromatograph and mass spectrophotometric analysis, to the NIH Tetramer Core Facility within four weeks of request approval. The address of the NIH Tetramer Core Facility is:
MHC Tetramer Core Facility
Emory University Vaccine Center
954 Gatewood Road
Atlanta, GA 30329
Phone: (404) 727-7215
Assume costs of shipping the tetramer reagents by providing FedEx or other carrier shipping account number (or by making arrangements for prepaid shipments) to the NIH Tetramer Core Facility. Tetramer reagents must be shipped at 4 oC. Monomeric soluble MHC/peptide must be shipped on dry ice (dry ice is provided by the NIH Tetramer Facility).
Complete and submit the original, signed Registration Form found on this site to the NIH Tetramer Facility administrative assistant at ThermoFisher BioServices. This form serves as the Materials Transfer Agreement (MTA) and must be completed to obtain tetramers.
Tetramer reagents are shipped with several package inserts including a form to confirm receipt of the reagent (this must be completed), a feedback form, a protocol for tetramer use and storage, and information regarding tetramer use restrictions.
Feedback forms are necessary for evaluating the quality of tetramer reagents produced at the NIH Tetramer Core Facility, as well as any technical problems or discoveries encountered by users. Therefore, all recipients of tetramers produced by the NIH Tetramer Core Facility must complete the Acknowledgement of Tetramer(s) Received and Status Report of Tetramer Use forms.
In all publications of studies utilizing tetramers supplied by the NIH Tetramer Facility, investigators are required to acknowledge the NIH Tetramer Facility as the source of the reagent(s). Copies of publications using tetramers from the NIH Tetramer Facility should be forwarded to the NIH Tetramer Resource Committee at the following address: NIAID Tetramer Facility, 6610 Rockledge Drive, Room 3007, Bethesda, MD 20892.
General inquires and information about pending requests should be sent to Dr. Alison Deckhut Augustine at 301-496-7551 or augustine@niaid.nih.gov. Technical inquiries or information about approved requests should be sent to Dr. Amy Stout at (404)-727-8117 or akstout@emory.edu.
