TCR Libraries
The present invention relates to a library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs may consist essentially of TCRs which may comprise an alpha chain variable domain from a natural repertoire and a beta chain variable domain from a natural repertoire, wherein the alpha chain variable domain may comprise a TRAV12-2 or a TRAV21 gene product and the beta chain variable domain may comprise a TRBV6 gene product.
1 . A library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs consists essentially of TCRs comprising an alpha chain comprising an alpha chain variable domain from a natural repertoire and a beta chain comprising a beta chain variable domain from a natural repertoire, wherein the alpha chain variable domain comprises a TRAV12-2 or a TRAV21 gene product and the beta chain variable domain comprises a TRBV6 gene product.
2 . A library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs consists essentially of TCRs comprising an alpha chain comprising an alpha chain variable domain from a natural repertoire and a beta chain comprising a beta chain variable domain from a natural repertoire, wherein the alpha chain variable domain comprises a TRAV12-2 or a TRAV21 gene product and the beta chain variable domain comprises a TRBV6 gene product and wherein at least a portion of the TCRs comprise an alpha chain variable domain and/or a beta chain variable domain comprising a non-natural mutation.
3 . The library according to claim 1 or claim 2 , wherein the TRBV6 gene product is a TRBV6-1, a TRBV6-2, a TRBV6-3, a TRBV6-5 or a TRBV6-6 gene product.
4 . The library according to claim 1 or claim 2 , wherein the TCR alpha chain variable domain comprises a TRAV12-2 gene product.
5 . The library according to claim 1 or claim 2 , wherein the TCR alpha chain variable domain comprises a TRAV21 gene product.
6 . The library according to claim 1 or claim 2 , wherein the alpha chain variable domain and the beta chain variable domain are displayed as a single polypeptide chain.
7 . The library according to claim 1 or claim 2 wherein the TCRs comprise a non-native disulphide bond between a constant region of the alpha chain and a constant region of the beta chain.
8 . The library according to claim 1 or claim 2 wherein the TCRs comprise a native disulphide bond between a constant region of the alpha chain and a constant region of the beta chain.
9 . The library according to claim 1 or claim 2 , wherein each alpha chain and each beta chain comprises a dimerization domain.
10 . The library according to claim 9 , wherein the dimerization domain is heterologous.
11 . The library according to claim 1 or claim 2 wherein the particles are phage particles.
12 . The library according to claim 1 or claim 2 wherein the particles are ribosomes.
13 . The library according to claim 1 or claim 2 wherein the particles are yeast cells.
14 . The library according to claim 1 or claim 2 wherein the particles are mammalian cells.
15 . A non-natural isolated T cell receptor (TCR) comprising a TCR alpha chain variable domain comprising a TRAV12-2 gene product or a TRAV21 gene product and a TCR beta chain variable domain comprising a TRBV6 gene product obtained from a library according to claim 1 or claim 2 .
16 . The TCR according to claim 15 , wherein the TRBV6 gene product is a TRBV6-1, a TRBV6-2, a TRBV6-3, a TRBV6-5 or a TRBV6-6 gene product.
17 . The TCR according to claim 15 , wherein the TCR is soluble.
18 . A method of identifying a TCR that specifically binds to a peptide antigen comprising screening the library according to claim 1 or claim 2 with the peptide antigen.
19 . A method of obtaining a T cell receptor that specifically binds a peptide antigen, comprising screening the library according to the first aspect of the invention with the peptide antigen, the method comprising:
a) panning the library using as a target the peptide antigen;
b) repeating step a) one or more times;
c) screening the phage clones identified in step a) or b); and
d) identifying a TCR that specifically binds the peptide antigen.
20 . A nucleic acid encoding a TCR alpha chain variable domain and/or a beta chain variable domain of the TCR according to claim 15 .
21 . A method of making a library of particles, the library displaying a plurality of different TCRs, the method comprising:
i) obtaining a plurality of nucleic acids that encode different TRAV12-2 or TRAV21 alpha chain variable domains;
ii) obtaining a plurality of nucleic acids that encode different TRBV6 beta chain variable domains;
iii) cloning the TRAV12-2 or TRAV21 alpha chain variable domain encoding nucleic acids into expression vectors;
iv) cloning the TRBV6 beta chain variable domain encoding nucleic acids into the same or different vectors; and
v) expressing the vectors in particles, thereby generating a library consisting essentially of TCRs comprising an alpha chain variable domain and a beta chain variable domain encoded by the nucleic acids.
22 . A method of making a library of particles, the library displaying a plurality of different TCRs, the method comprising:
i) obtaining a plurality of nucleic acids that encode different TRAV12-2 or TRAV21 alpha chain variable domains using primers that hybridise to nucleic acids encoding TRA12-2 or TRAV21 alpha chain variable domains;
ii) obtaining a plurality of nucleic acids that encode different TRBV6 beta chain variable domains using primers that hybridise to nucleic acids encoding TRAV6 beta chain variable domains;
iii) cloning the TRAV12-2 or TRAV21 alpha chain variable domain encoding nucleic acids into expression vectors;
iv) cloning the TRBV6 beta chain variable domain encoding nucleic acids into the same or different vectors; and
v) expressing the vectors in particles, thereby generating a library consisting essentially of TCRs comprising an alpha chain variable domain and a beta chain variable domain encoded by the nucleic acids to which said primers hybridise.
23 . The method of claim 21 or claim 22 , wherein the nucleic acids of step (i) and step (ii) are obtained from a natural repertoire.
24 . The method of claim 21 or claim 22 , comprising a further step of introducing non-natural mutations to the nucleic acids.
25 . The method of claim 21 or claim 22 , wherein non-natural mutations are introduced to the nucleic acids prior to step iii).
26 . The method according to claim 21 or claim 22 , wherein the TRBV6 nucleic acid sequence is a TRBV6-1, a TRBV6-2, a TRBV6-3, a TRBV6-5 or a TRBV6-6 gene product.
27 . A method according to claim 21 or claim 22 , wherein the TCR alpha chain variable domain and the TCR beta chain variable domain are expressed as a single chain polypeptide.
28 . The method of obtaining a T cell receptor that specifically binds a peptide antigen, comprising screening the library according to claim 1 or claim 2 with the peptide antigen.
29 . A particle displaying on its surface a TCR according to claim 15 .
30 . The particle according to claim 29 , wherein the particle is a phage particle, a ribosome, a yeast cell or a mammalian cell.