Method of producing pharmaceutical compositions comprising immunogenic chikungunya virus chikv-delta5nsp3

A pharmaceutical composition comprising

• (i) CHIKV-Δ5nsP3 particles comprising a genome as defined by the polynucleotide sequence of SEQ ID NO:
• 1;
• (ii) CHIKV-Δ5nsP3 particles having genome that differs from SEQ ID NO:
• 1 by having sequence heterogeneity at nucleic acid positions 8543, 8882, 9119 and/or 9649 corresponding to G55R, E168K, E247K and A423A (silent) point mutations in the E2 structural protein according to SEQ ID NO:
• 2;
• and
• (iii) optionally a pharmaceutically acceptable excipient,
• wherein 1-50% of the CHIKV-Δ5nsP3 particles in the composition having a genome with heterogeneity at nucleic acid 8543, 8882, 9119 and/or 9649 compared with SEQ ID NO:
• 1.

A pharmaceutical composition comprising

• (i) CHIKV-Δ5nsP3 particles having a genome as defined by the polynucleotide sequence of SEQ ID NO:
• 1;
• and
• (ii) CHIKV-Δ5nsP3 particles having a genome with sequence heterogeneity at one or more nucleic acid positions in the E2 protein coding region of SEQ ID NO:
• 1, wherein said genomic sequence heterogeneity results in G55R, G82R, H99Y, E168K, T230I, H232Y, E247K and/or E247D mutations with respect to the E2 structural protein according to SEQ ID NO:
• 2;
• and
• (iii) optionally a pharmaceutically acceptable excipient,
• wherein 1-50% of the CHIKV-Δ5nsP3 particles in the composition having a genome with sequence heterogeneity resulting in G55R, G82R, H99Y, E168K, T230I, H232Y, E247K and/or E247D mutations with respect to the E2 structural protein according to SEQ ID NO:
• 2.