US 11,698,375 B2
Method for producing an examination reagent and kit for analysing a T-cell frequency
Sebastian Hartmut Springer, Bremen (DE); and Sunil Kumar Saini, Kerala (IN)
Assigned to CONSTRUCTOR UNIVERSITY BREMEN GGMBH, Bremen (DE)
Filed by CONSTRUCTOR UNIVERSITY BREMEN GGMBH, Bremen (DE)
Filed on Jul. 1, 2019, as Appl. No. 16/458,203.
Application 16/458,203 is a continuation of application No. 14/370,217, granted, now 10,386,367, previously published as PCT/DE2013/100001, filed on Jan. 3, 2013.
Claims priority of application No. 20 2012 100 019.9 (DE), filed on Jan. 4, 2012; and application No. 10 2012 101 028.5 (DE), filed on Feb. 8, 2012.
Prior Publication US 2019/0331681 A1, Oct. 31, 2019
Int. Cl. G01N 33/566 (2006.01); C07K 14/74 (2006.01); G01N 33/569 (2006.01)
CPC G01N 33/566 (2013.01) [C07K 14/70539 (2013.01); G01N 33/56977 (2013.01); G01N 2333/70539 (2013.01)] 8 Claims
 
1. A method for producing an examination reagent, the method comprising:
providing and adding a helper ligand peptide to an unfolded MHC class I receptor protein and β2m in order to produce a folded MHC class I receptor protein, wherein the helper ligand peptide consists of a peptide of 8 to 11 amino acid residues in length and has a lower affinity for a folded MHC class I receptor protein than that of an examination peptide of interest;
adding a glycyl-leucine (GL) dipeptide as a ligand exchanger/ligand releaser to the folded MHC class I receptor protein, wherein the GL dipeptide has a lower affinity for the folded MHC class I receptor protein than the affinity of the helper ligand peptide for the folded MHC class I receptor protein; and
adding an examination peptide having a higher affinity for the folded MHC class I receptor protein than the affinity of the helper ligand peptide for the folded MHC class I receptor protein, so that the examination peptide is exchanged for the helper ligand peptide, thereby providing the examination reagent, and wherein the MHC class I receptor protein is H-2Kb.