Methods and compositions for recombinant dengue viruses or vaccine and diagnostic development

A chimeric dengue virus E glycoprotein, comprising the following amino acid substitutions wherein the numbering is based on the reference amino acid sequence of an E glycoprotein of dengue virus serotype 3 (DENV3) identified as SEQ ID NO:

• 1:
• (i) Q46L, A50V, L53P, T55V, T138S, Q156H, V158T, T159I, E161T, S167P, T169S, A171I, I172Q, P174T, E175D, T178A, N270T, G273T, S275T, D382E, and N383K;
• (ii) Q46L, T138S, I141V, Q156H, V158T, T159I, E161T, S167P, T169S, A171I, I172Q, P174T, E175D, T178A, G180T, E182D, N270T, G273T, S275T, E291T, N302G, T303S, V305K, E323K, K325E, E327T, N375Y, I378V, I380A, D382E, N383K, I387L;
• or
• (iii) Q46L, Q52N, L53P, T138S, I141V, Q156H, V158T, T159I, E161T, S167P, T169S, A171I, I172Q, P174T, E175D, T178A, G180T, E182D, N270T, G273T, S275T, E291T, N302G, T303S, V305K, E323K, K325E, E327T, N375Y, I378V, I380A, D382E, N383K, I387L, and N388S,
• and wherein said dengue virus E glycoprotein further comprises an insertion of the amino acid residues T and E between amino acid residues 155 and 156.

A chimeric dengue virus E glycoprotein, comprising the following amino acid substitutions wherein the numbering is based on the reference amino acid sequence of an E glycoprotein of dengue virus serotype 1 (DENV1) identified as SEQ ID NO:

• 2:
• (i) L46Q, N52Q, S138T, T156Q, E157S, T160V, I161T, T163E, S171T, I173A, Q174I, T176P, D177E, T272N, T275G, T277S, E384D, and K385N;
• (ii) L46Q, S138T, V141I, T156Q, T160V, I161T, T163E, P169S, S171T, I173A, Q174I, T176P, D177E, A180T, T182G, D184E, T272N, T275G, T277S, and T293E, and wherein said dengue virus E glycoprotein further comprises a deletion of the amino acid residues E157 and H158;
• (iii) P53L, V55T, A63G, S66T, T68I, T81V, V83P, T88Q, F90Y, R93K, R94H, F96Y, I114V, K120Q, V122L, T123E, K124P, L125I, I129V, V197I, K203N, S205A, L207M, K210R, L214F, S225T, S227E, Q228T, E229P, Q234K, D235E, T242N, and T272N;
• (iv) A63G, S66T, T68I, T81V, V83P, T88Q, F90Y, R93K, R94H, F96Y, I114V, K120Q, V122L, T123E, K124P, L125I, L214F, S225T, S227E, Q228T, E229P, Q234K, D235E, T242N, and T272N;
• (v) L46Q, V50A, N52Q, P53L, V55T, A63G, S66T, T68I, T81V, V83P, T88Q, F90Y, R93K, R94H, F96Y, I114V, K120Q, V122L, T123E, K124P, L125I, I129V, V197I, K203N, S205A, L207M, K210R, L214F, S225T, S227E, Q228T, E229P, Q234K, D235E, T242N, T272N, T275G, and T277S;
• (vi) V50A, N52Q, P53L, V55T, A63G, S66T, T68I, T81V, V83P, T88Q, F90Y, R93K, R94H, F96Y, I114V, K120Q, V122L, T123E, K124P, L125I, I129V, V197I, K203N, S205A, L207M, K210R, L214F, S225T, S227E, Q228T, E229P, Q234K, D23SE, T242N, T272N, T275G, and T277S;
• (vii) V300A, G304N, S305T, K307V, E309K, A313S, V320I, V322I, Q323K, K325E, E327K, T329E, S339T, Q340E, V345K, Q347H, I357V, D360K, K363E, Y377N, V380I, A382I, E384D, K385N, L389I, and S390N;
• (viii) V300A, G304N, S305T, K307V, E309K, A313S, V320I, V322I, Q323K, K325E, E327K, T329E, S339T, Q340E, V345K, T346A, Q347H, I357V, D360K, K363E, Y377N, V380I, A382I, E384D, K385N, L389I, and S390N;
• or
• (ix) V300A, G304N, S305T, K307V, E309K, A313S, V320I, V322I, Q323K, K325E, E327K, T329E, S339T, Q340E, E342G, K343Q, V345K, T346A, Q347H, I357V, D360K, K363E, Y377N, V380I, A382I, E384D, K385N, L389I, and S390N.