Compositions and methods for immunotherapy
The present invention provides compositions and methods for immunotherapy, which include shelf-stable pharmaceutical compositions for inducing antigen-specific T cells. Such compositions are employed as components of an artificial antigen presenting cell (aAPC), to provide a patient with complexes for presentation of an antigen (e.g., a tumor antigen) and/or a T cell co-stimulatory molecule.
1. An anti-CD28 antibody comprising
an immunoglobulin heavy chain variable region with the amino acid sequence selected from:
EVKLQQSGPGLVKPSETLSLTCTVSGFSLSDYGVHWVRQAPGKGLEWLG VIWAGGGTNYNSALMSRKTISKDNSKSQVFLKMNSLTAADTAVY YCARDKGYSYYYSMDYWGQGTLVTVSS (SEQ ID NO: 2), or
EVKLQQSGPGLVKPSETLSLTCTVSGFSLSDYGVHWVRQAPGKGLEWLG VIWAGGGTNYNSALMSRKTISKDNSKSQVSLKMSSVTAADTAVY YCARDKGYSYYYSMDYWGQGTLVTVSS (SEQ ID NO: 4),
and an immunoglobulin light chain variable region with the amino acid sequence selected from:
DIELTQSPDSLAVSLGERATINCRASESVEYYVTSLMQWYQQKPGQPPKL LIFAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAMYFCQQSR KVPYTFGGGTKVEIK (SEQ ID NO: 8).
2. A pharmaceutical composition comprising a polymeric bead or particle, an antibody according to claim 1 , and an antigen presenting complex comprising a humanized immunoglobulin heavy chain sequence fused to an HLA amino acid sequence, wherein the complex optionally does not contain an immunoglobulin light chain sequence.
3. The anti-CD28 antibody of claim 1 , wherein the antibody comprises an immunoglobulin heavy chain with the amino acid sequence selected from:
EVKLQQSGPGLVKPSETLSLTCTVSGFSLSDYGVHWVRQAPGKGLEWLG VIWAGGGTNYNSALMFLSRKTISKDNSKSQVKMNSLTAADTAVY YCARDKGYSYYYSMDYWGQGTLVTVSSASTKGPSVFPLAPCSRS TheSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSL SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKGLPSSIEKTISKAláKGQPREPQVYTLPPSQEEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS RLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLCLSLGK (SEQ ID NO: 22), or
EVKLQQSGPGLVKPSETLSLTCTVSGFSLSDYGVHWVRQAPGKGLEWLG VIWAGGGTNYNSALMSRKTISKDNSKSQVSLKMSSVTAADTAVY YCARDKGYSYYYSMDYWGQGTLVTVSSASTKGPSVFPLAPCSRS TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQF NWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVS LTCLVKGFYPSDIAVETWESNGQPENNYKTTPPVLDSDGSFFLYSR LTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLCLSLGK (SEQ ID NO: 21);
and an immunoglobulin light chain with the amino acid sequence selected from:
IELTQSPDSLAVSLGERATINCRASESVEYYVTSLMQWYQQKPGQPPKLLI FAASNVESGVPDRFSGSGSGTDTLTISSLQAEDVAMYFCQQSRKVP YTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20).
4. The anti-CD28 antibody of claim 1 , wherein the antibody comprises a constant region, and the constant region comprises one or more mutations suitable for chemical coupling the antibody to a solid support.
5. The anti-CD28 antibody of claim 4 , wherein the constant region is IgG4 isotype, and optionally comprises one or more hinge stabilizing mutations.
6. The anti-CD28 antibody of claim 4 , wherein the antibody comprises an unpaired cysteine coupled to a solid support.
7. The anti-CD28 antibody of claim of claim 5 , wherein the antibody comprises an unpaired cysteine at S473C.
8. The anti-CD28 antibody of claim 6 , wherein the solid support is a bead or particle.
9. The antibody of claim 4 , wherein the constant region further comprises one or more mutations to reduce Fc gamma receptor binding.
10. The antibody of claim 9 , wherein the one or more mutations that reduce Fc gamma receptor binding comprise L248E of an IgG4 sequence.
11. The anti-CD28 antibody of claim 8 , wherein the bead or particle comprises a polymer or block co-polymer.
12. The anti-CD28 antibody of claim 8 , wherein the bead or particle further comprises molecular complexes presenting antigen for recognition by T cells.
13. The anti-CD28 antibody of claim 12 , wherein the molecular complex presenting antigen comprises MHC Class I and/or MHC Class II complexes.
14. The antibody of claim 12 , wherein the molecular complex is an HLA-Ig fusion complex.