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Identifizierung von Antigen-Epitopen

Identifying T cell epitopes in protein antigens, useful e.g. for preparing vaccines and for isolating specific T cells, uses nanoparticles as carriers for a receptor in a competitive binding assay.
 
: Brunner, H.; Tovar, G.; Schiestel, T.; Mueller, C.; Flad, T.

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Frontpage ()

DE 2003-10310261 A: 20030305
DE 2003-10310261 A: 20030305
WO 2004-EP2170 A: 20040303
DE 10310261 A1: 20040923
G01N0033
C12N0001
German
Patent, Electronic Publication
Fraunhofer IGB ()

Abstract
Die vorliegende Erfindung betrifft ein Verfahren zur Identifizierung und/oder zum Nachweis von T-Zell-Epitopen eines Protein-Antigens, Verfahren zur Herstellung von Peptid-Impfstoffen gegen ein Protein-Antigen, Verfahren zur Qualitaetskontrolle von Rezeptor-Ligand-Komplexen und/oder deren Bestandteilen, Verfahren zur Herstellung von Nanopartikeln mit mindestens einer immobilisierten Rezeptor-Einheit oder einem immobilisierten Rezeptor, Verfahren zur Herstellung von Nanopartikeln mit immobilisierten Rezeptor-Ligand- Komplexen, insbesondere Peptid-praesentierenden MHC-Molekuelen, Verfahren zur Anreicherung und/oder Isolierung spezifischer CD4+-T- oder CD8+-T-Lymphozyten aus peripheren mononuklearen Blutzellen, Verfahren zum Primen einer CD8+-T-Lymphozyten-Reaktion in vitro, Nanopartikel mit einer immobilisierten Rezeptor-Einheit, insbesondere einer immobilisierten Kette eines MHC-Molekuels, Nanopartikel mit einem immobilisierten Rezeptor, insbesondere einem immobilisierten MHC-Molekuel, Nanopartikel mit einem immobilisierten Rezeptor-Ligand-Komplex, insbesodnere einem Peptid-praesentierenden MHC-Molekuel, einen Peptid-Impfstoff, einen Kit zur Identifizierung und/oder zum Nachweis von T-Zell-Epitopen eines Protein-Antigens, sowie die Verwendung der Nanopartikel zur Identifizierung und/oder zum Nachweis von T-Zell-Epitopen, zur Herstellung von Peptid-Impfstoffen, zur Anreicherung und/oder Isolierung spezifischer T-Lymphozyten sowie zum Primen einer ...

 

WO2004078909 A UPAB: 20041015 NOVELTY - In vitro method for identification and/or detection of the T cell epitopes (A) of a protein antigen (Ag). DETAILED DESCRIPTION - In vitro method for identification and/or detection of the T cell epitopes (A) of a protein antigen (Ag) in which: (a) a population of peptide fragments (F) from Ag is subjected to competitive binding assay to a first, immobilized receptor subunit (SU1), preferably in presence of a second receptor subunit (SU2) which, together with SU1, forms a receptor; (b) at least one (F) with affinity for at least SU1, preferably also SU2, binds and (c) the bound (F) is isolated and analyzed. The new feature is that SU1 having at least one functional group (FG1) is immobilized on a nanoparticle (NP) having a functional group (FG2) reactive with FG1 so that a receptor-(F) complex is formed on NP, and this, or the bound (F), is analyzed. INDEPENDENT CLAIMS are also included for the following: (1) method for identifying and/or preparing peptide vaccines against Ag, using (A) identified by the new method; (2) method for quality control of receptor-ligand complexes and/or their components; (3) method for preparing NP having a receptor, or at least one of its subunits, immobilized on the surface; (4) method for preparing NP having a peptide-presenting MHC (major histocompatibility complex) molecule immobilized on it; (5) method for enrichment and/or isolation of specific CD4+ or CD8+ T cells from peripheral blood mononuclear cells; (6) method for priming and/or restimulating a CD4+ or CD8+ T lymphocyte reaction in vitro; (7) NP that contain at least one receptor subunit, especially a chain of an MHC molecule, immobilized on it; (8) NP with an immobilized MHC molecule on its surface; (9) peptide vaccine that comprises a peptide-presenting MHC molecule, obtained by method (1) and/or an (A) identified by the new method; and (10) kit for identification or detection of (A). ACTIVITY - None given. MECHANISM OF ACTION - Vaccine. No relevant biological data given. USE - T cell epitopes (A), identified by the new method and optionally formulated as a complex with an MHC molecule, are used in preparation of vaccines against Ag; for enrichment or isolation of specific CD4+/CD8+ T cells from peripheral blood (for subsequent expansion and return to the donor) and for priming or restimulating such cells in vitro. ADVANTAGE - The method provides simultaneous and rapid examination of many peptide sequences.

: http://publica.fraunhofer.de/documents/N-24803.html