FLEXamers: A Double Tag for Universal Generation of Versatile Peptide-MHC Multimers

• Published in: The Journal of immunology (1950) Vol. 202; no. 7; pp. 2164 - 2171
• Main Authors: Effenberger, Manuel, Stengl, Andreas, Schober, Kilian, Gerget, Maria, Kampick, Maximilian, Müller, Thomas R, Schumacher, Dominik, Helma, Jonas, Leonhardt, Heinrich, Busch, Dirk H
• Format: Journal Article
• Language: English
• Published: United States 01.04.2019
• Subjects: Animals; Cell Separation - methods; Histocompatibility Antigens - analysis; Humans; Receptors, Antigen, T-Cell - analysis; T-Lymphocytes - immunology
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.1801435

Peptide-MHC (pMHC) multimers have become a valuable tool for immunological research, clinical immune monitoring, and immunotherapeutic applications. Biotinylated tetramers, reversible Streptamers, or dye-conjugated pMHC multimers are distinct pMHC reagents tailored for T cell identification, traceless T cell isolation, or TCR characterization, respectively. The specific applicability of each pMHC-based reagent is made possible either through conjugation of probes or reversible multimerization in separate production processes, which is laborious, time-consuming, and prone to variability between the different types of pMHC reagents. This prohibits broad implementation of different types of pMHC reagents as a standard toolbox in routine clinical immune monitoring and immunotherapy. In this article, we describe a novel method for fast and standardized generation of any pMHC multimer reagent from a single precursor ("FLEXamer"). FLEXamers unite reversible multimerization and versatile probe conjugation through a novel double tag (Strep-tag for reversibility and Tub-tag for versatile probe conjugation). We demonstrate that FLEXamers can substitute conventional pMHC reagents in all state-of-the-art applications, considerably accelerating and standardizing production without sacrificing functional performance. Although FLEXamers significantly aid the applicability of pMHC-based reagents in routine workflows, the double tag also provides a universal tool for the investigation of transient molecular interactions in general.