Modified chimeric coronavirus spike protein for enhancement of viral titers
AssigneeiosBio Ltd
InventorJeffrey DREW
Life of this patent
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Abstract
The disclosure relates to the production of viral particles, in particular adenoviral particles for use in vaccines. Modified chimeric coronavirus spike antigens are disclosed, together with polynucleotides encoding said antigens, viral particles and methods of production/formulation of the viral particles.
Claims32
A polynucleotide encoding an antigen from a virus, wherein the antigen has been modified (i) to target the antigen to the plasma membrane and/or (ii) to reduce membrane fusion and/or the formation of syncytia.
The polynucleotide of any one of the preceding claims, wherein the cytoplasmic tail is replaced with the cytoplasmic tail from infectious bronchitis virus in which the dilysine motif is substituted, such as wherein the dilysine motif is substituted with alanine, optionally wherein:
- (a) the tyrosine internalisation sequence is also mutated to replace the tyrosines, optionally with alanine;
- and/or
- (b) the cytoplasmic tail comprises one or more additional amino acid substitutions, deletions or insertions.
The polynucleotide of any one of the preceding claims, wherein the modifications to reduce membrane fusion and/or the formation of syncytia comprise the substitution of tryptophan residues, optionally wherein the tryptophan residues are in a transmembrane domain of the antigen, such as in an aromatic stretch of the transmembrane domain.
The polynucleotide of any one of the preceding claims, wherein the antigen is from a coronavirus, such as wherein the antigen is the spike (S) protein of SARS-CoV-2.
The polynucleotide of any one of the preceding claims, which encodes a SARS-CoV-2 spike protein comprising:
- (a) the following sequence, or encodes a spike protein with at least 90% identity to the following sequence FVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQD LFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWI FGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESE FRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKH TPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTA GAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKG IYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVA DYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQ TGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF ERDISTEIYQAGSTPCNGVEGENCYFPLQSYGFQPTNGVGYQPYRVVVLSFE LLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFG RDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCT EVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGI CASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVT TEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQD KNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTL ADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLA GTITSGWTFGAGAALQIPFAMQMAYRENGIGVTQNVLYENQKLIANQFNS AIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDIL SRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECV LGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICH DGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVN NTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRL NEVAKNLNESLIDLQELGKYKTYIKAPAYVALAIAFLTIIFILILGWVFFMTG CCGCCCGCFGIIPLMSKCSKKSSAATTFDNDVVTEQYRPAASV (SEQ ID NO:
- 17), or (b) the sequence as shown in FIG. 1 or with at least 90% identity to the sequence shown in FIG. 1.
Dependencies are inferred from the claim text.