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- Title
Increased yields and biological potency of knob-into-hole-based soluble MHC class II molecules.
- Authors
Serra, Pau; Garabatos, Nahir; Singha, Santiswarup; Fandos, César; Garnica, Josep; Solé, Patricia; Parras, Daniel; Yamanouchi, Jun; Blanco, Jesús; Tort, Meritxell; Ortega, Mireia; Yang, Yang; Ellestad, Kristofor K.; Santamaria, Pere
- Abstract
Assembly of soluble peptide-major histocompatibility complex class II (pMHCII) monomers into multimeric structures enables the detection of antigen-specific CD4+ T cells in biological samples and, in some configurations, their reprogramming in vivo. Unfortunately, current MHCII-αβ chain heterodimerization strategies are typically associated with low production yields and require the use of foreign affinity tags for purification, precluding therapeutic applications in humans. Here, we show that fusion of peptide-tethered or empty MHCII-αβ chains to the IgG1-Fc mutated to form knob-into-hole structures results in the assembly of highly stable pMHCII monomers. This design enables the expression and rapid purification of challenging pMHCII types at high yields without the need for leucine zippers and purification affinity tags. Importantly, this design increases the antigen-receptor signaling potency of multimerized derivatives useful for therapeutic applications and facilitates the detection and amplification of low-avidity T cell specificities in biological samples using flow cytometry. Recombinant MHC class II molecules are instrumental in antigen-specific T-cell identification assays and showed efficacy as experimental medicines. Here, the authors engineer MHC class II molecules with species-specific knob-into-hole heteromerization domains, enabling a translatable purification process with improved stability, yields, and biological potency.
- Subjects
MAJOR histocompatibility complex; MONOMERS; T cells; IMMUNOGLOBULIN G; CELLULAR signal transduction
- Publication
Nature Communications, 2019, Vol 10, Issue 1, pN.PAG
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-019-12902-2