US 11,732,026 B2
Glycoprotein hormone receptor mutations
Bernard Rees Smith, Cardiff (GB); Jadwiga Furmaniak, Cardiff (GB); Jane Sanders, Barry (GB); and Jennifer Miller-Gallacher, Cardiff (GB)
Assigned to RSR Limited, Cardiff (GB)
Appl. No. 15/317,687
Filed by RSR Limited, Cardiff (GB)
PCT Filed Jun. 11, 2015, PCT No. PCT/GB2015/000171
§ 371(c)(1), (2) Date Dec. 9, 2016,
PCT Pub. No. WO2015/189543, PCT Pub. Date Dec. 17, 2015.
Claims priority of application No. 1410409 (GB), filed on Jun. 11, 2014.
Prior Publication US 2017/0204159 A1, Jul. 20, 2017
Int. Cl. C07K 14/705 (2006.01); C07K 14/72 (2006.01); G01N 33/68 (2006.01)
CPC C07K 14/723 (2013.01) [G01N 33/6854 (2013.01)] 8 Claims
 
1. A mutant thyroid stimulating hormone receptor (TSHR) which is a mutated form of human wild type TSHR shown in SEQ ID NO: 2 or fragment thereof, which mutant TSHR or fragment thereof is able to bind TSHR autoantibodies and comprises residues 22-260 of the TSHR as shown in SEQ ID NO: 4 or a sequence having 9011% sequence identity therewith, wherein the mutant TSHR or fragment thereof, has increased thermostability with respect to the equivalent non-mutated wild type TSHR or fragment thereof, wherein thermostability refers to the half-life of the mutant TSHR or fragment thereof, as compared to the half-life of the equivalent wild-type TSHR or fragment thereof, as measured under identical conditions in a binding assay which determines the amount of mutant TSHR or fragment thereof that retains ability to bind, at the test temperature, an autoantibody selected from the group consisting of: M22, KI-70; K1-18, and patient serum autoantibodies, wherein the half-life of the mutant TSHR or fragment thereof at 42° C. is 1.5 times greater or more than the half-life of the equivalent wild type TSHR or fragment thereof, and wherein the mutant TSHR or fragment thereof comprises one or more mutations are within residues 22 to 260 of the mutant TSHR as shown in SEQ ID NO: 2 or fragment thereof, wherein the one or more mutations are selected from the group consisting of:
1) I253R;
2) D143P and I253R;
3) R112P and D143P and I253R;
4) R112P and D143P and D151E and I253R;
5) R112P and D143P and D151E and V169R and I253R;
6) H63C and R112P and D143P and D151E and V169R and I253R;
7) H63C and R112P and D143P and V169R and I253R; and
8) H63C and R112P and D143P and S166T and I253R.