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A monoclonal antibody that recognizes a unique 13-residue epitope in the cytoplasmic tail of HLA-E.
Verhaar, Elisha R; Gan, Jin; Buhl, Susan; Li, Ziao; Horowitz, Amir; Ploegh, Hidde L.
Afiliación
  • Verhaar ER; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, the Netherlands.
  • Gan J; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
  • Buhl S; Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, NY 10461, USA.
  • Li Z; Department of Oncological Sciences, Precision Immunology Institute, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
  • Horowitz A; Department of Oncological Sciences, Precision Immunology Institute, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
  • Ploegh HL; Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, the Netherlands. Electronic address: hidde.ploegh@childrens.harvard.edu.
Mol Immunol ; 172: 56-67, 2024 Aug.
Article en En | MEDLINE | ID: mdl-38901180
ABSTRACT
The Class I MHC molecule (MHC-I) HLA-E presents peptides that are derived from the signal sequences, either those of other MHC-I products, or of viral type I membrane glycoproteins. Monoclonal antibodies with proven specificity for HLA-E, and with no cross-reactions with other MHC-I products, have yet to be described. To obtain anti-HLA-E-specific antibodies suitable for a range of applications, we generated monoclonal antibodies against a unique feature of HLA-E its cytoplasmic tail. We created an immunogen by performing an enzymatically catalyzed transpeptidation reaction to obtain a fusion of the cytoplasmic tail of HLA-E with a nanobody that recognizes murine Class II MHC (MHC-II) products. We obtained a mouse monoclonal antibody that recognizes a 13-residue stretch in the HLA-E cytoplasmic tail. We cloned the genes that encode this antibody in expression vectors to place an LPETG sortase recognition motif at the C-terminus of the heavy and light chains. This arrangement allows the site-specific installation of fluorophores or biotin at these C-termini. The resulting immunoglobulin preparations, labeled with 4 equivalents of a fluorescent or biotinylated payload of choice, can then be used for direct immunofluorescence or detection of the tag by fluorescence or by streptavidin-based methods. We also show that the 13-residue sequence can serve as an epitope tag, independent of the site of its placement within a protein's sequence. The antibody can be used diagnostically to stain for HLA-E on patient tumor samples, it can be used as an antibody-epitope tag for extracellular proteins, and it enables research into the unique role of the cytoplasmic tail of HLA-E.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Antígenos de Histocompatibilidad Clase I / Antígenos HLA-E / Anticuerpos Monoclonales / Epítopos Límite: Animals / Humans Idioma: En Revista: Mol Immunol Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Antígenos de Histocompatibilidad Clase I / Antígenos HLA-E / Anticuerpos Monoclonales / Epítopos Límite: Animals / Humans Idioma: En Revista: Mol Immunol Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido