Viral glycoprotein

Abstract

There is provided a viral glycoprotein comprising a glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein and / or a modified N-terminal that allows the glycoprotein to be resistant towards detect and / or degradation. The mutations may include lysine to arginine mutations at residues 491, 493, 496, 497 and 511 of the VSV glycoprotein. Also disclosed in a polynucleotide, a vector, a host cell, a composition, a method of treating a disease, and use of the glycoprotein in the manufacture of a medicament for treating a disease.

Description

[0001] VIRAL GLYCOPROTEIN

[0002] TECHNICAL FIELD

[0003] The present disclosure relates broadly to an alternative viral glycoprotein for pseudotyping with viruses to improve virus titer and infectivity of target cells.

[0004] BACKGROUND

[0005] Viral vectors are widely used for research, therapeutic, and diagnostic applications due to their ability to efficiently deliver genetic material into target cells. Viral glycoproteins / envelope glycoproteins mediate host cell recognition, binding, and entry. Accordingly, viral glycoproteins / envelopes are recognized as critical determinants of vector tropism, stability and production efficiency.

[0006] Pseudotyping viral vectors with viral glycoproteins have been employed to expand the range of targetable cell types, enhance transduction efficiency and improve particle stability. However, many viral glycoproteins are susceptible to intracellular degradation or misfolding in producer cells, which can limit viral titers and reduce overall vector performance.

[0007] Strategies to modify or engineer viral glycoproteins to enhance target cell binding and improve stability in production cells are therefore desirable to optimize viral vector design and production.

[0008] Therefore, there is a need to provide an alternative viral glycoprotein.

[0009] SUMMARY

[0010] In one aspect, there is provided a viral glycoprotein comprising a glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein and / or a modified N-terminal that allows the glycoprotein to be resistant towards detection and / or degradation.

[0011] In some examples, the one or more mutations is a substitution at the C-terminal or at positions 491 to 511.

[0012] In some examples, the one or more mutations is a substitution at position 491, 493, 496, 497, 511, or combinations thereof.In some examples, the one or mutations is a substitution to an amino acid with a positively charged side chain and / or an amino acid with a polar uncharged side chain.

[0013] In some examples, the one or more mutations comprise one or more substitutions to an amino acid that is not subjected to ubiquitylation / degradation.

[0014] In some examples, the one or more mutations comprise one or more substitutions to an arginine (“R”) and / or to an asparagine (“N”).

[0015] In some examples, the modified N-terminal comprises a binding domain that binds to an immune cell.

[0016] In some examples, the glycoprotein comprises the sequence:

[0017] VSVG Ctrl (pMDLG)_amino acid (SEQ ID NO: 1) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgk

[0018] > VSVG1R_amino acid (SEQ ID NO: 2) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRlkh tkkrqiytdiemnrlgk

[0019] > VSVG2R_amino acid (SEQ ID NO: 3) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciklRh tkkrqiytdiemnrlgk

[0020] > VSVG1-2R_amino acid (SEQ ID NO: 4) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htkkrqiytdiemnrlgk

[0021] > VSVG3,4R_amino acid (SEQ ID NO: 5) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht RRrqiytdiemnrlgk

[0022] > VSVG1-4R_amino acid (SEQ ID NO: 6) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgk

[0023] > VSVG5R_amino acid (SEQ ID NO: 7) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgR

[0024] > VSVG1-5R_amino acid (SEQ ID NO: 8) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgR,

[0025] > Affi-VSVGwt / Affi-VSVG (SEQ ID NO: 19) Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvlrvgihlciklkhtkkrqiytdiemnrlgk,

[0026] > Affi-VSVG1-5R (SEQ ID NO: 24) mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvIrvgihlciRIRhtRRrqiytdiemnrlgR

[0027] or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0028] In some examples, the glycoprotein is encoded by a sequence comprising > VSVG Ctrl (pMDLG)_nt (SEQ ID NO: 9) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0029] > VSVG1R_nt (SEQ ID NO: 10) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggatattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtgaagca caccaagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0030] > VSVG2R_nt (SEQ ID NO: 11) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattctggttctccgagttggtatccatctttgcattaaattGaGAcaca cAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag> VSVG1-2R_nt (SEQ ID NO: 12) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtGaGAc acacAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0031] > VSVG3,4R_nt (SEQ ID NO: 13) atgaagtgcctttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caGACGCagacagatttatacagacatagagatgaaccgacttggaaag

[0032] > VSVG1-4R_nt (SEQ ID NO: 14) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatagagatgaaccgacttggaaag

[0033] > VSVG5R_nt (SEQ ID NO: 15) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggatattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatCgagatgaaccgGctGggCaGA

[0034] > VSVG1-5R_nt (SEQ ID NO: 16) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattctggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatCgagatgaaccgGctGggCaGA> Affi-VS Gwt_nt (SEQ ID NO: 17) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tatatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcataaattaaa gcacaccaagaaaagacagatttatacagacatagagatgaaccgacttggaaagtaa

[0035] > Affi-VSVG1-5R_nt (SEQ ID NO: 18) atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgccatgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtG aGAcacacAaGACGCagacagatttatacagacatagagatgaaccgGctGggCaGA,

[0036] or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above.

[0037] In some examples, the glycoprotein comprises a combination of glycoproteins selected from any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17 (Affi-VSVGwt) or SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); and

[0038] any one of the glycoproteins with sequences: SEQ ID NO: 2 (VSVG1R), SEQ ID NO: 3 (VSVG2R), SEQ ID NO: 4 (VSVG1-2R), SEQ ID NO: 5 (VSVG3,4R), SEQ ID NO:6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R), or SEQ ID NO: 1 (VSVG (wild type / control)).

[0039] In some examples, the glycoprotein comprises a combination of glycoproteins selected from any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17 (Affi-VSVGwt) or SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); and any one of the glycoproteins with sequences: SEQ ID NO: 5 (VSVG3,4R), SEQ ID NO: 6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R), or SEQ ID NO: 1 (VSVG (wild type / control)).

[0040] In some examples, the glycoprotein comprises the combination of glycoproteins with sequences: SEQ ID NO: 10 (VSVG1-5R) and SEQ ID NO: 18 (Affi-VSVG1-5R).

[0041] In another aspect, there is provided a polynucleotide encoding any one of the glycoproteins as disclosed herein.

[0042] In yet another aspect, there is provided a vector encoding the polynucleotide or glycoprotein as disclosed herein.

[0043] In yet another aspect, there is provided a host cell comprising the polynucleotide, the vector, or glycoprotein as disclosed herein.

[0044] In yet another aspect, there is provided a composition comprising the glycoprotein or polynucleotide as disclosed herein.

[0045] In some examples, the glycoprotein as disclosed herein is for use in therapy, optionally wherein the therapy comprises gene therapy.

[0046] In yet another aspect, there is provided a method of treating a disease, comprising providing the glycoprotein as disclosed herein.

[0047] In yet another aspect, there is provided a use of the glycoprotein as disclosed herein in the manufacture of a medicament for treating a disease, optionally wherein the medicament is a gene therapy.

[0048] DEFINITIONS

[0049] The term “mutation” as used herein refers to a change in the nucleic acid sequence of the genome of an organism.

[0050] The term “substitution” as used herein refers to a type of mutation in which one nucleotide is replaced by a different nucleotide. The term can also refer to the replacement of one amino acid in a protein with a different amino acid.The term “point mutation” as used herein refers to a mutation where a single nucleotide base is changed, inserted or deleted from a nucleic acid (such as deoxyribose nucleic acid (DNA), messenger ribonucleic acid (mRNA)) sequence of an organism’s genome. A point mutation in a protein sequence refers to a change in one amino acid of the protein that results from a point mutation in the underlying nucleic acid such as DNA or mRNA i.e., a point mutation at the nucleotide level changes a codon. If that codon now specifies a different amino acid, the protein sequence changes at exactly one position. A point mutation occurs at one specific point in the genome. Point mutations can be grouped into two main categories i) base substitution and ii) base insertion I deletion. Examples of base substitution may include silent mutation where nucleotide changes but the amino acid does not change; missense mutation where nucleotide changes and causes a different amino acid which may affect protein function; nonsense mutation which is a mutation that produces a stop codon and leads to premature termination of the protein. Examples of base insertion or deletion may include frameshift mutation that occurs when insertion / deletion is not in multiples of three and therefore shifting the reading frame and usually producing a non-functional protein.

[0051] The term “substitution” and “point mutation” is used interchangeably herein. The term “truncation” as used herein refers to a shortened gene or shortened transcript typically caused by deletions, early stop codons, splicing defects, and the like. A protein truncation refers to a protein that is shorter than the original sequence because of a mutation that creates a nonsense mutation, a frameshift mutation, or cleavage or degradation that removes C-terminal or N-terminal segments. A protein that lacks part of its normal sequence may lose function or misfold.

[0052] The term “modified protein / modification of a protein” as used herein refers to any changes made to a protein compared to its original / wild type form. This may include sequence modification (such as point mutation, substitution, insertion, deletion, truncation, fusion proteins, and the like), post-translational modification (PTM) which include chemical modifications added after translation (such as phosphorylation, glycosylation, ubiquitination, acetylation, methylation, sumoylation, proteolytic cleavage, and the like) or artificial or engineered modifications to change or enhance protein functions (such as His-tag for purification, stabilizing mutations to improve folding or expression, engineered glycosylation patterns, and the like).

[0053] The term “binding domain” as used herein refers to a structurally and functionally distinct region of a molecule that mediates specific binding to a target partner such as DNA or another protein.The term “pseudotyping” as used herein refers to the process of producing a viral particle that contains the core and genome of one virus but displays the envelope or surface glycoprotein of a different virus, thereby altering its cell entry properties.

[0054] The term “VSV-G or affVSV-G pseudotyped virus” as used herein refers to a viral vector whose core and genome core comes from one virus (such as lentivirus, retrovirus, and the like) but whose surface envelope protein is the VSV-G. VSV-G pseudotyping allows broad tropism i.e., VSV-G binds widely expressed receptors, allowing entry into many cell types. VSV-G pseudotyping confers strong physical ability enabling virus concentration by ultracentrifugation. In addition, VSV-G pseudotyping provides high transduction efficiency.

[0055] The term “chimeric / chimera” as used herein refers to something that is made by combining parts from two or more different biological sources into a single entity.

[0056] The term "associated with", used herein when referring to two elements refers to a broad relationship between the two elements. In genomics, “associated” typically refers to entities that are found in a correlative relationship (e.g., when A is upregulated / downregulated, B is also upregulated I downregulated). “Associated” is used when one cannot establish the causal or functional relationship based on correlation.

[0057] The term "and / or", e.g., "X and / or Y" is understood to mean either "X and Y" or "X or Y" and should be taken to provide explicit support for both meanings or for either meaning.

[0058] Further, in the description herein, the word “substantially” whenever used is understood to include, but not restricted to, "entirely" or “completely” and the like. In addition, terms such as "comprising", "comprise", and the like whenever used, are intended to be non-restricting descriptive language in that they broadly include elements / components recited after such terms, in addition to other components not explicitly recited. For example, when “comprising” is used, reference to a “one” feature is also intended to be a reference to “at least one” of that feature. Terms such as “consisting”, “consist”, and the like, may in the appropriate context, be considered as a subset of terms such as "comprising", "comprise", and the like. Therefore, in embodiments disclosed herein using the terms such as "comprising", "comprise", and the like, it will be appreciated that these embodiments provide teaching for corresponding embodiments using terms such as “consisting”, “consist”, and the like. Further, terms such as "about", "approximately" and the like whenever used, typically means a reasonable variation, for example a variation of + / - 5% of the disclosed value, or avariance of 4% of the disclosed value, or a variance of 3% of the disclosed value, a variance of 2% of the disclosed value or a variance of 1% of the disclosed value.

[0059] The phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” “having,” “containing,” “involving,” and variations thereof, is meant to encompass the items listed thereafter and additional items. Use of ordinal terms such as “first,” “second,” “third,” etc., in the claims to modify a claim element does not by itself connote any priority, precedence, or order of one claim element over another or the temporal order in which acts of a method are performed. Ordinal terms are used merely as labels to distinguish one claim element having a certain name from another element having a same name (but for use of the ordinal term), to distinguish the claim elements. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the inventions embodied therein herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention.

[0060] The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0061] Throughout this disclosure, certain embodiments may be disclosed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.Other embodiments are within the following claims and non-limiting examples. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. Exemplary embodiments of the present invention are provided in the following examples. While the exemplary embodiments are described in sufficient detail to enable those skilled in the art to practice the invention, it should be understood that the present invention is not limited to these examples.

[0062] Furthermore, in the description herein, certain values may be disclosed in a range. The values showing the end points of a range are intended to illustrate a preferred range. Whenever a range has been described, it is intended that the range covers and teaches all possible sub-ranges as well as individual numerical values within that range. That is, the end points of a range should not be interpreted as inflexible limitations. For example, a description of a range of 1% to 5% is intended to have specifically disclosed sub-ranges 1% to 2%, 1% to 3%, 1% to 4%, 2% to 3% etc., as well as individually, values within that range such as 1%, 2%, 3%, 4% and 5%. It is to be appreciated that the individual numerical values within the range also include integers, fractions and decimals. Furthermore, whenever a range has been described, it is also intended that the range covers and teaches values of up to 2 additional decimal places or significant figures (where appropriate) from the shown numerical end points. For example, a description of a range of 1% to 5% is intended to have specifically disclosed the ranges 1.00% to 5.00% and also 1.0% to 5.0% and all their intermediate values (such as 1.01 %, 1.02% ... 4.98%, 4.99%, 5.00% and 1.1%, 1.2% ... 4.8%, 4.9%, 5.0% etc.,) spanning the ranges. The intention of the above specific disclosure is applicable to any depth / breadth of a range.

[0063] Additionally, when describing some embodiments, the disclosure may have disclosed a method and / or process as a particular sequence of steps. However, unless otherwise required, it will be appreciated that the method or process should not be limited to the particular sequence of steps disclosed. Other sequences of steps may be possible. The particular order of the steps disclosed herein should not be construed as undue limitations. Unless otherwise required, a method and / or process disclosed herein should not be limited to the steps being carried out in the order written. The sequence of steps may be varied and still remain within the scope of the disclosure.

[0064] Furthermore, it will be appreciated that while the present disclosure provides embodiments having one or more of the features / characteristics discussed herein, oneor more of these features / characteristics may also be disclaimed in other alternative embodiments and the present disclosure provides support for such disclaimers and these associated alternative embodiments.

[0065] DESCRIPTION OF EMBODIMENTS

[0066] The present disclosure addresses the lack of binding of viruses to target cells, and the limitations imposed on virus production by the production cells.

[0067] In one aspect, there is provided a viral glycoprotein comprising a glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein and / or a modified N-terminal that allows the glycoprotein to be resistant towards detection and / or degradation.

[0068] In one aspect, there is provided a viral glycoprotein comprising a glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein that allows the glycoprotein to be resistant towards detection and / or degradation.

[0069] In some examples, there is provided a viral glycoprotein comprising a viral glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein.

[0070] In some examples, the viral glycoprotein is an envelope protein.

[0071] In some examples, the viral glycoprotein is Vesicular stomatitis virus glycoprotein (VS -G).

[0072] In some examples, the VSV-G glycoprotein may include the sequence / amino acid sequence:

[0073] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgk (SEQ ID NO: 1) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0074] VSV-G is a viral glycoprotein used across the life science industry for the pseudotyping of viruses (such as lentivirus and retrovirus) used in cell and gene therapyprocesses due to its broad tropism allowing the VSV-G pseudotyped virus to infect almost any cell line used in research or manufacturing. VSV-G uses only one envelope protein for receptor binding and membrane fusion therefore allowing pseudotyping to be straightforward compared to viruses that need multiple envelope proteins.

[0075] In some examples, the viral glycoprotein I VSV-G of the present disclosure is used for the pseudotyping of enveloped viruses / viral vectors. In some examples, the enveloped viruses / viral vectors may include but are not limited to lentivirus, retrovirus, alpha and flavivirus, adeno-associated virus (AAV), rabies virus, measles virus, and the like.

[0076] In some examples, the viral glycoprotein / VSV-G pseudotyped viruses can transduce / infect cells expressing receptors / entry factors such as but are not limited to low-density liporeceptor receptor (LDR) family, phosphatidylserine-mediated uptake heparan sulfate proteoglycans, and the like.

[0077] In some examples, the cells expressing receptors I entry factors for the viral glycoprotein / VSV-G pseudotyped viruses may include but are not limited to somatic cells expressing LDLR family receptors (such as epithelial cells, fibroblasts, neurons, hepatocytes), immune cells expressing LDLR at lower levels (such as T cells, B cells, natural killer cells), macrophages, dendritic cells, endothelial cells (which allow entry via for example phosphatidylserine pathway), and the like.

[0078] These viruses pseudotyped with VSV-G can transduce / infect cells expressing for example the cognate receptor of the VSV-G protein, the Low-Density Lipoprotein -Receptor (LDL-R). The LDL-R is a ubiquitous protein expressed by a majority of cells across the human body but with major differences in the level of expression from one cell type to another. Certain immune cells (such as natural killer cells (NK cells), B cells, virus specific T cells (VSTs) express LDL-R only transiently and at very low levels. Therefore, these cells become extremely difficult to transduce with standardized viruses pseudotyped with VSV-G. In addition, cellular viral immunity can target viral gene products, including VSV-G, for degradation. VSV-G is also susceptible to degradation during viral production, storage and concentration therefore leading to lower viral titers.

[0079] The inventors of the present disclosure have generated VSV-G viral glycoprotein mutants (VSVGmut) that led to an increased production of mature 3rd generation lentiviral particles when used in place of the standard VSV-G glycoprotein. The present disclosure provides degradation resistant chimeric VSV-G for enhanced viral production.A mutated version of the VSV-G gene (VSVGmut) is introduced into the plasmid encoding the VSV-G protein, which is normally transfected into packaging cells during the lentivirus production process. The plasmid replaces the VSV-G plasmid during lentivirus production.

[0080] Alternatively, the VSVGmut coding sequence can be stably introduced into a packaging cell line, for constitutive or inducible production of the VSVGmut during the lentivirus production process. VSVGmut again replaces VSV-G in such cell lines. Specific residues have been mutated to prevent the recognition of VSV-Gmut by cellular degradation proteins and thus preventing their destruction. Resistance to cellular degradation allows more viral and chimeric glycoproteins to be expressed incorporated into viral envelopes, generating a higher number of intact viruses, thus increasing the viral titer. High titer is critical for cell and gene therapy manufacturing and can affect the cost or even the feasibility of a prospective therapy.

[0081] In some examples, the one or more mutations may include a substitution / point mutation.

[0082] In some examples, the one or more mutations are at the C-terminal. In some examples, the one or more mutations are at the intracellular / cytoplasmic domain.

[0083] In some examples, the one or more mutations at the C-terminal / intracellular domain I cytoplasmic domain produces glycoprotein / viral glycoprotein that are resistant to the cellular degradation machinery (VSVGmut). In some examples, the one or more mutations at the C-terminal / intracellular domain / cytoplasmic domain produces a degradation resistant glycoprotein / viral glycoprotein / envelope protein.

[0084] VSV-G is a type I transmembrane glycoprotein / viral glycoprotein with an extracellular N-terminus, transmembrane domain that anchors the protein in the membrane and C-terminal intracellular / cytoplasmic domain. The intracellular / cytoplasmic domain of VSV-G plays a key role in protein trafficking, stability and incorporation into virions which indirectly affects resistance to degradation and detection.

[0085] In some examples, the one or more mutations may include an amino acid substitution at the C terminal. In some examples, the glycoprotein may include 1, or 2, or 3, or 4, or 5 amino acid mutations / substitutions thereof, and the like. In some examples, the glycoprotein may include one or more, two or more, three or more, four or more, or five or more amino acid mutations / substitutions. In some examples, the glycoprotein may include one amino acid mutation / substitution. In some examples, the glycoprotein may include two amino acid mutations / substitutions. In some examples, the glycoprotein may include three amino acid mutations / substitutions. In someexamples, the glycoprotein may include four amino acid mutations / substitutions. In some examples, the glycoprotein may include five amino acid mutations / substitutions.

[0086] In some examples, the one or more mutations is a substitution at the C-terminal or at positions 491 to 511. In some examples, the one or more amino acid mutations may include amino acid substitutions at positions 491 to 511. In some examples, the one or more amino acid mutations may include an amino acid substitution at the C-terminal or at positions 491 to 511. In some examples, the one or more amino acid mutations may include a point mutation at the C-terminal or at positions 491 to 511.

[0087] In some examples, the one or more amino acid mutations / substitution may be a substitution at the C-terminal at positions 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, and / or 511. In some examples, the one or more amino acid mutations I substitution may be a substitution / point mutation at the C-terminal or at positions 491, 492, 493, 494, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, and / or 511. In some examples, the one or more mutations may include a substitution at position 491, 493, 496, 497, 511, or combinations thereof. In some examples, the one or more amino acid mutations may include a substitution at position 491, 493, 496, 497, 511, or combinations thereof. In some examples, the one or more amino acid mutations may include a point mutation at position 491 , 493, 496, 497, 511, or combinations thereof.

[0088] In some examples, the glycoprotein may include one or more, two or more, three or more, four or more, or five or more amino acid mutations / substitutions at position 491, 493, 496, 497, 511, or combinations thereof. In some examples, the glycoprotein may include at least one, at least two, at least three, at least four, at least five amino acid mutations / substitutions. In some examples, the glycoprotein may include at least one, at least two, at least three, at least four, or at least five amino acid mutations / substitutions at position 491, 493, 496, 497, 511, or combinations thereof. In some examples, the glycoprotein may include up to five amino acid mutations I substitutions. In some examples, the glycoprotein may include up to five amino acid mutations / substitutions at position 491, 493, 496, 497, 511, or combinations thereof.

[0089] In some examples, the amino acid mutations may include mutations of 1, or 2, or 3, or 4, or 5 lysine residues. In some examples, the glycoprotein may include one or more, two or more, three or more, four or more, or five or more mutations / substitutions in lysine residues. In some examples, the glycoprotein may include one amino acid mutation / substitution in a lysine residue. In some examples, the glycoprotein may include two amino acid mutations / substitutions in lysine residues. In some examples,the glycoprotein may include three amino acid mutations / substitutions in lysine residues. In some examples, the glycoprotein may include four amino acid mutations / substitutions in lysine residues. In some examples, the glycoprotein may include five amino acid mutations / substitutions in lysine residues.

[0090] In some examples, the one or more mutations may include a substitution to an amino acid that is not subjected to ubiquitylation / degradation.

[0091] In some examples, the one or more amino acid mutations may include a substitution to an amino acid with a positively charged side chain and / or an amino acid with a polar uncharged side chain. In some examples, the one or more amino acid mutations may include a substitution to an amino acid with a positively charged side chain and / or an amino acid with a polar uncharged side chain.

[0092] In some examples, the one or more mutations may include one or more substitutions to an arginine (“R”) and / or to an asparagine (“N”). In some examples, the one or more amino acid mutations may include one or more substitutions to an arginine (“R”) and / or to an asparagine (“N”).

[0093] In some examples, the one or more amino acid mutations may include a substitution to an amino acid with a positively charged side chain.

[0094] In some examples, the one or more amino acid mutations may include a substitution to an arginine (“R”), or a histidine (“H”).

[0095] In some examples, the amino acid mutation may include a substitution to an arginine (“R”).

[0096] In some examples, various nucleotide substitution combinations may be used to obtain the same amino acid substitutions as disclosed herein. In some examples, the corresponding number of nucleotide substitutions required to effect the amino acid substitutions as disclosed herein may vary, as an amino acid (such as arginine, histidine, lysine) may be encoded by multiple alternative codons. Accordingly, the number of nucleotide substitutions depends on the specific codon choices employed.

[0097] In some examples, the glycoprotein may comprise a amino acid substitution / point mutation at position 491. In some examples, the amino acid substitution I point mutation is K491R.

[0098] In some examples, the VSVG glycoprotein with the K491R substitution / point mutation (VSVG1R) may include the amino acid sequence:

[0099] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRlkh tkkrqiytdiemnrlgk (SEQ ID NO: 2) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0100] In some examples, the VSVG glycoprotein with the K491R substitution / point mutation (VSVG1R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtgaagca caccaagaaaagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 10) or a polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0101] In some examples, the glycoprotein may comprise a substitution / point mutation at position 493. In some examples, the substitution / point mutation is K493R.

[0102] In some examples, the VSVG glycoprotein with K493R substitution / point mutation (VSVG2R) may include the amino acid sequence:Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciklRh tkkrqiytdiemnrlgk (SEQ ID NO: 3) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0103] In some examples, the VSVG glycoprotein with K493R substitution I point mutation (VSVG2R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattGaGAcaca cAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 11) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.In some examples, the glycoprotein may comprise two substitutions / point mutations at positions 491 and 493. In some examples, the two substitutions I point mutation are K491R and K493R.

[0104] In some examples, the VSVG glycoprotein with K491R and K493R substitutions / point mutations (VSVG1-2R) may include the amino acid sequence:

[0105] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htkkrqiytdiemnrlgk (SEQ ID NO: 4) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0106] In some examples, the VSVG glycoprotein with K491R and K493R substitutions / point mutations (VSVG1-2R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtGaGAcacacAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 12) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0107] In some examples, the glycoprotein may comprise two substitutions / point mutations at positions 496 and 497. In some examples, the two substitutions I point mutation is K496R and K497R.

[0108] In some examples, the VSVG glycoprotein with K496R and K497R substitutions / point mutations (VSVG3.4R) may include the amino acid sequence:

[0109] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgw mchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvde ytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkack mqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispv dlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfply mighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihl ciklkhtRRrqiytdiemnrlgk (SEQ ID NO: 5) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0110] In some examples, the VSVG glycoprotein with K496R and K497R substitutions / point mutations (VSVG3.4R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caGACGCagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 13) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0111] In some examples, the glycoprotein may comprise four substitutions / point mutations at positions 491, 493, 496 and 497. In some examples, the four substitutions / point mutations are K491R, K493R, K496R and K497R.

[0112] In some examples, the VSVG glycoprotein with K491R, K493R, K496R and K497R substitutions / point mutations (VSVG1-4R) may include the amino acid sequence:

[0113] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgk (SEQ ID NO: 6) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0114] In some examples, the VSVG glycoprotein with K491R, K493R, K496R and K497R substitutions / point mutations (VSVG1-4R) may include the nucleotide sequence:

[0115] Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattctggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 14) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0116] In some examples, the glycoprotein may comprise a substitution I point mutation at position 511. In some examples, the substitution / point mutation is K511 R.

[0117] In some examples, the VSVG glycoprotein with K511R substitutions / point mutations (VSVG5R) may include the amino acid sequence:

[0118] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgR (SEQ ID NO: 7) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0119] In some examples, the VSVG glycoprotein with K511R substitutions / point mutations (VSVG5R) may include the nucleotide sequence:

[0120] Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatagagatgaaccgGctGggCaGA (SEQ ID NO: 15) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0121] In some examples, the glycoprotein may include five substitutions I point mutations at positions 491, 493, 496, 497 and 511. In some examples, the five substitutions / point mutations are K491R, K493R, K496R, K497R and K511R.

[0122] In some examples, the VSVG glycoprotein with K491 , K493R, K496R, K497R and K511R substitutions I point mutations (VSVG1-5R) may include the amino acid sequence:

[0123] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgR (SEQ ID NO: 8) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0124] In some examples, the VSVG glycoprotein with K491R, K493R, K496R, K497R and K511R substitutions / point mutations (VSVG1-5R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactacttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttatttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatagagatgaaccgGctGggCaGA (SEQ ID NO: 16) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0125] In some examples, the one or mutation is a substitution to an amino acid with a polar uncharged side chain.

[0126] In some examples, the one or more mutation is a substitution to an asparagine (“N”), a serine (“S”), a threonine (“T”), or a glutamine (“Q”).

[0127] In some examples, the mutation is a substitution to an asparagine (“N”).

[0128] In some examples, the five amino acid substitutions / point mutations are K491 N, K493N, K496N, K497N and K511N.

[0129] In some examples, the VSVG glycoprotein with K491N, K493N, K496N, K497N and K511N substitutions I point mutations (VSVG1-5N) may include the amino acid sequence:

[0130] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciNIN htNNrqiytdiemnrlgN (SEQ ID NO: 20) or an amino acid sequence having at least 60%,70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0131] In some examples, the VSVG glycoprotein with K491N, K493N, K496N, K497N and K511N substitutions I point mutations (VSVG1-5N) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaCttaaaCcaca ccaaCaaCagacagatttatacagacatagagatgaaccgacttggaaaC (SEQ ID NO: 21) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0132] In some examples, the five amino acid substitutions / point mutations are K491 N, K493R, K496N, K497R and K511N.

[0133] In some examples, the VSVG glycoprotein with K491N, K493R, K496N, K497R and K511 N substitutions / point mutations (VSVG-NRNRN) may include the amino acid sequence:

[0134] Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciNIR htNRrqiytdiemnrlgN (SEQ ID NO: 22) or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0135] In some examples, the VSVG glycoprotein with K491N, K493R, K496N, K497R and K511N substitutions / point mutations (VSVG-NRNRN) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaCttaCGgcaca ccaaCCGCagacagatttatacagacatagagatgaaccgacttggaaaC (SEQ ID NO: 23) or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0136] In some examples, there is provided a glycoprotein / viral glycoprotein comprising a viral glycoprotein comprising one or more mutations not at / within the intracellular domain of the glycoprotein. In some examples, the one or more mutations are at the extracellular domain / non-cytoplasmic domain. In some examples, the glycoprotein / viral glycoprotein may include a glycoprotein where the glycoprotein is not able I cannot bind to its target / target cell. In some examples, the VSV-G glycoprotein that is not able I cannot bind to its target / target cell is a “blinded VSV-G glycoprotein / glycoprotein mutant”. In some examples, the VSV-G glycoprotein that are not able / cannot bind to its target / target cell may include one or more mutations that are not in the intracellular / cytoplasmic domain.

[0137] In some examples, the one or more mutations may comprise one or more substitutions / point mutations at positions 8, 47, 354, or combinations thereof. In some examples, the one or more substitutions / point mutations at positions 8, 47, 354, or combinations are not within the intracellular domain.

[0138] In some examples, the glycoprotein may further comprise one or more substitutions I point mutations of H8A, K47Q, R354A, or combinations thereof. In some examples, the one or more substitutions / point mutations of H8A, K47Q, R354A, or combinations thereof are not within the intracellular domain.

[0139] In some examples, the glycoprotein may comprise a truncation mutation.

[0140] In some examples, the glycoprotein may comprise a truncation at the N-terminal. In some examples, the glycoprotein may further comprise a truncation of the first 420 amino acid residues. In some examples, the truncation mutation / the truncation at the N-terminal is not at / within the intracellular domain. For example, where the glycoprotein is a variant of VSV-G protein, the glycoprotein may comprise a truncation of the first 420 amino acids of the VSV-G.

[0141] The inventors of the present disclosure have generated a VSVG-binding domain fusion protein (Affi-VSVG) that led to increased infectivity of the binding domain’s target expressing cells.

[0142] A binding domain is fused to the extracellular portion of VSV-G. This allows the Affi-VSVG pseudotyped virus to bind to cells with low binding capacity for wildtype VSV-G. An important cell type with low VSV-G binding capacity are T cells, which are used in many cell therapies.

[0143] In some examples, the modified N-terminal may include a binding domain that binds to an immune cell.

[0144] In some examples, the glycoprotein may comprise a modified N-terminal comprising a binding domain. In some examples, the binding domain is an affinity binding domain.

[0145] In some examples, the binding domain may be a single-chain variable fragment (scFv) of an antibody; an affibody; a nanobody; a DARPIN; a ligand; a cytokine; a VHH,and the like. In some examples, the binding domain is an antibody. In some examples, the binding domain is a single chain antibody. In some examples, the binding domain is single chain antibody targeting an immune cell. In some examples, the binding domain is a T cell single chain antibody / a single chain antibody that targets a T cell. In some examples, the binding domain is a pan T cell single chain antibody. In some examples, the binding domain is a pan T cell single chain antibody which binds to a protein on the T cell surface. In some examples, the binding domain is a pan T cell single chain antibody which binds to the CD3 protein on the immune cell / T cell surface.

[0146] Advantageously, the glycoprotein with the binding domain of the present application produced infectious viral titers comparable to the glycoprotein alone, targeting CD3 positive immune cells.

[0147] In some examples, the binding domain binds to an immune cell. In some examples, the binding domain / modified N-terminal allows the glycoprotein to bind to a cellular target other than its physiological ligand, which are low / transiently expressed in cells / immune cells relevant for cell therapies such as T cells.

[0148] In some examples, the immune cell may include, but is not limited to, a lymphocyte (such as B cells and T cells), an NK cell, a neutrophil, a monocyte, a dendritic cell, a macrophage, an antigen presenting cell, and the like. In some examples, the lymphocyte may be a B lymphocyte (i.e. B cell), a T lymphocyte (i.e. T cell), and the like. In some examples, the T lymphocyte may be a CD4+ T cell, a CD8+ T cell, and the like. In some examples, the T cells may be virus-specific T cells (VSTs). In some examples, the present disclosure includes transferring of a CAR gene into T cells to generate CAR-T cells I therapeutic CAR-T cells.

[0149] In some examples, the VSVG glycoprotein with binding domain (Affi-VSVGwt / Affi-VSVG) may include the amino acid sequence:

[0150] Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvlrvgihlciklkhtkkrqiytdiemnrlgk (SEQ ID NO: 19) or an amino acidsequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0151] In some examples, the VSVG glycoprotein with binding domain (Affi-VSVGwt / Affi-VSVG) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaa gcacaccaagaaaagacagatttatacagacatagagatgaaccgacttggaaag (SEQ ID NO: 17) or apolynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to the sequence.

[0152] In some examples, the mutations to generate VSVGmut were combined with the structural layout of the chimeric VSV-G protein to produce Affi-VSVGmut. In some examples, the glycoprotein / viral glycoprotein may include an N terminal chimera / modified N-terminal chimera and with one or more mutations in the intracellular domain / cytoplasmic domain.

[0153] The inventors of the present disclosure then combined the mutated and fusion VSV-G protein (Affi-VSVGmut). Lentivirus produced using the Affi-VSVGmut possesses higher titer generally and higher infectivity of target cell types compared to lentivirus produced with a wild-type VSV-G.

[0154] The chimeric Affi-VSVG is mutated similarly to VSVGmut, to protect it from cellular degradation. The virus pseudotyped with Affi-VSVGmut can be produced at higher titer than those produced with Affi-VSVG and transduce its target cell types more efficiently than VSV-G or VSVGmut.

[0155] High titer lentivirus produced using the VSV-Gmut alone or Affi-VSVGmut can be used to transduce target cell types in vitro and in vivo.

[0156] In some examples, the VS VG glycoprotein with a N terminal chimera / modified N-terminal chimera and with K491R, K493R, K496R, K497R and K511R substitutions (Affi-VSVG 1-5R) may include the amino acid sequence:

[0157] Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgv qpedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhn dligtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqsc gyatvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgel sslgkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqd verildysicqetwskiragipispvdisylapknpgtgpaftiingtikyfetryirvdiaapilsrmvgmisgttterelwddw apyedveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielveg wfsswkssiasfffiigliiglfMrvgihlciRIRhtRRrqiytdiemnrlgR (SEQ ID NO: 20)

[0158] In some examples, the VSVG glycoprotein with a N terminal chimera / modified N-terminal chimera and with K491R, K493R, K496R, K497R and K511R substitutions (Affi-VSVG 1-5R) may include the nucleotide sequence: atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgccatgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtG aGAcacacAaGACGCagacagatttatacagacatagagatgaaccgGctGggCaGA (SEQ ID NO: 18),

[0159] or a polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above.

[0160] The inventors of the present disclosure have engineered the VSV-glycoprotein in two distinct fashions:

[0161] i) by addition of a binding domain including but not limited to a single-chain variable fragment (scFv) of an antibody; an affibody; a nanobody; a DARPIN; a ligand; a VHH; tothe N-terminal side of the VSV-G protein leading to a chimeric and synthetic glycoprotein that can be used to envelope the retro- and lentiviral particles and / or

[0162] II) by substitution(s) I point mutation(s) of specific residues located in the intracellular domain of the VSV-G protein to become resistant towards the proteins involved in the detection and degradation of viral proteins.

[0163] In some examples, the present disclosure may include the combination of i) a glycoprotein with one or more mutations in the intracellular domain / cytoplasmic domain and ii) a glycoprotein with an N terminal chimera / modified N-terminal chimera and one or more mutations in the intracellular domain / cytoplasmic domain (i.e. , VSVGmut + Affi-VSVGmut).

[0164] In some examples, the glycoproteins may include a combination of glycoproteins selected any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17(Affi-VSVGwt), SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); and

[0165] any one of the glycoproteins with sequences: SEQ ID NO: 2 (VSVG1R), SEQ ID NOL 3 (VSVG2R), SEQ ID NO: 4 (VSVG1-2R), SEQ ID NO: 5 (VSVG3,4R), SEQ ID NO: 6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R), or SEQ ID NO: 1 (VSVG (wild type / control)).

[0166] In some examples, the glycoproteins may include a combination of glycoproteins selected from any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17 (Affi-VSVGwt), SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); and any one of the glycoproteins with sequences: SEQ ID NO: 5 (VSVG3,4R), SEQ ID NO: 6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R), or SEQ ID NO: 1 (VSVG (wild type / control))

[0167] In some examples, the glycoproteins may include the combination of glycoproteins with sequence SEQ ID NO: 8 (VSVG1-5R) and SEQ ID NO: 17 (AffiVSVGwt) In some examples, the present disclosure may include the combination of glycoproteins with the sequence SEQ ID NO: 8 (VSVG1-5R) and SEQ ID NO: 18 (Affi-VSVG1-5R). Advantageously the combination of glycoproteins with sequence SEQ ID NO: 8 (VSVG-1-5R) and SEQ ID NO: 18 (Affi-VSVG1-5R) yield the highest titer in viral production.

[0168] In some examples, the glycoprotein comprises the amino acid sequence:

[0169] VSVG Ctrl (pMDLG / pALD) amino acid (SEQ ID NO: 1)Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgk

[0170] > VSVG1R_amino acid (SEQ ID NO: 2) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRlkh tkkrqiytdiemnrlgk

[0171] > VSVG2R_amino acid (SEQ ID NO: 3) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciklRh tkkrqiytdiemnrlgk

[0172] > VSVG1-2R_amino acid (SEQ ID NO: 4) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htkkrqiytdiemnrlgk> VSVG3,4R_amino acid (SEQ ID NO: 5) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht RRrqiytdiemnrlgk

[0173] > VSVG1-4R_amino acid (SEQ ID NO: 6) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgk

[0174] > VSVG5R_amino acid (SEQ ID NO: 7) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgR

[0175] > VSVG1-5R_amino acid (SEQ ID NO: 8) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgR,> VSVG1-5N (SEQ ID NO: 20) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciNIN htNNrqiytdiemnrlgN,

[0176] > Affi-VSVGwt / Affi-VSVG (SEQ ID NO: 19) Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvlrvgihlciklkhtkkrqiytdiemnrlgk,

[0177] > Affi-VSVG 1-5R (SEQ ID NO: 24) mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvIrvgihlciRIRhtRRrqiytdiemnrlgR

[0178] or an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above,or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

[0179] In some examples, the glycoprotein is encoded by a nucleotide sequence comprising:

[0180] > VSVG Ctrl (pMDLG / pALD)_nt (SEQ ID NO: 9) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0181] > VSVG1R_nt (SEQ ID NO: 10) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtgaagca caccaagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0182] > VSVG2R_nt (SEQ ID NO: 11) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggatattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattGaGAcaca cAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0183] > VSVG1-2R_nt (SEQ ID NO: 12) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtGaGAc acacAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag

[0184] > VSVG3,4R_nt (SEQ ID NO: 13) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggatattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caGACGCagacagatttatacagacatagagatgaaccgacttggaaag

[0185] > VSVG1-4R_nt (SEQ ID NO: 14) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatagagatgaaccgacttggaaag

[0186] > VSVG5R_nt (SEQ ID NO: 15) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatCgagatgaaccgGctGggCaGA

[0187] > VSVG1-5R_nt (SEQ ID NO: 16) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggatattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatCgagatgaaccgGctGggCaGA

[0188] > VSVG1-5N_nt (SEQ ID NO: 21) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaCttaaaCcaca ccaaCaaCagacagatttatacagacatagagatgaaccgacttggaaaC

[0189] > Affi-VSVGwt_nt (SEQ ID NO: 17) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgatcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaa gcacaccaagaaaagacagatttatacagacatagagatgaaccgacttggaaagtaa

[0190] > Affi-VSVG1-5R_nt (SEQ ID NO: 18) atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgtggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtG aGAcacacAaGACGCagacagatCtaCacCgacatCgagatgaaccgGctGggCaGA, or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above.

[0191] In another aspect, there is provided a polynucleotide encoding any one of the glycoprotein as disclosed herein.

[0192] In yet another aspect, there is provided a vector encoding the polynucleotide or glycoprotein as disclosed herein.

[0193] In yet another aspect, there is provided a host cell comprising the polynucleotide, the vector, or glycoprotein as disclosed herein.

[0194] In yet another aspect, there is provided a composition comprising the glycoprotein or polynucleotide as disclosed herein.

[0195] In some examples, the glycoprotein as disclosed herein for use in a therapy, such as gene therapy. In some examples, the glycoprotein as disclosed herein for use in therapy, optionally wherein the therapy comprises gene therapy.

[0196] In yet another aspect, there is provided a method of treating a disease, comprising providing the glycoprotein as disclosed herein.

[0197] In yet another aspect, there is provided use of the glycoprotein as disclosed herein in the manufacture of a medicament for treating a disease, optionally wherein the medicament is a gene therapy.

[0198] In yet another aspect, there is provided use of the glycoprotein as disclosed herein in the manufacture of a medicament, such as a gene therapy.

[0199] In some examples, gene therapy may include cell therapy.

[0200] In some examples, there is provided a glycoprotein as described herein.

[0201] In some examples, there are methods or kits or reagents provided as disclosed herein.

[0202] RESULTSVSV-G, the envelope glycoprotein of the Vesicular Stomatitis Virus, is degraded by cellular antiviral protein MARCHF8 via its lysine residues. Mutation of the five intracellular lysine residues of VSV-G to arginine prevented the degradation of VSV-G in biochemical and virus production assays, resulting in higher virus production. Lentivirus pseudotyped with VSV-G mutant with five K to R mutations (VSVG1-5R) transduced a higher percentage of cells than the same with wildtype VSV-Gwt (FIG. 1).

[0203] VSV-G pseudotyped lentivirus are a standard tool to genetically modify cells. However, some cell types are inefficiently transduced by lentivirus, including primary T cells commonly used in cell therapy. Low, transient expression of the VSV-G ligand is thought to be a mechanism causing low efficiency. A molecule with affinity for some cell surface receptor on T cells, fused to the VSV-G protein, can increase binding of the virus to the target cells thereby increasing transduction efficiency or titer.

[0204] The inventors of the present disclosure fused the SP34 antibody, which recognizes CD3e on T cells, to the wild-type VSV-G. Lentivirus pseudotyped with the resulting Affi-VSVG transduced T cells more efficiently than VSV-G alone. T cells were more efficiently transduced across the first three days of T cell culture (FIG. 2), when LDLR, the VSV-G ligand, is reported to vary widely and have direct impact on lentiviral transduction efficiency.

[0205] The inventors of the present disclosure reasoned that the mutations which protected VSV-G from degradation would also protect Affi-VSVG. The inventors of the present disclosure mutated the five intracellular Lysine residues in Affi-VSVG to Arginine. Lysine mutated VSV-G combined with Affi-VSVGwt produced higher titers than VSVGwt and Affi-VSVGwt. Combining the Lysine mutated VSVG with Lysine mutated Affi-VSVG produced the highest amount of TalH T cells infectious titers (FIG. 3).

[0206] The inventors of the present disclosure found that mutating residues 496 and 497 provided partial protection from degradation and improved VSVG-pseudotyped and Affi-VSVG pseudotyped lentivirus titers (FIG. 4). Similarly, VSV-G containing four Lysine mutations but missing K511R provided partial protection and improved lentivirus titers.

[0207] Mutation of ubiquitylation targeted Lysine residues to Arginine often preserves protein function due to the chemical similarity between Lysine andArginine, with the latter being unsuitable for ubiquitylation. The inventors of the present disclosure noted that the intracellular Ks are highly conserved in wild-type VSV Indiana strains, but rare, individual K to R mutations exist, as do K to N (Asparagine). The inventors of the present disclosure replaced all K residues with N, or a combination of N and R.

[0208] Lentiviruses are a standard tool to introduce therapeutic genes to cells in ex vivo or in vivo settings. Lentiviruses are used to deliver the gene encoding for a Chimeric Antigen Receptor in approved ex-vivo CAR-T cell therapies and in ongoing in vivo delivery of CAR vectors clinical trials.

[0209] MATERIALS & METHODS

[0210] Cells and Culture Conditions

[0211] HEK293T (ATCC) or HEK293T-LentiX (TakaraBio) cells were used for lentivirus packaging. HEK cells were cultured in DM EM media supplemented with 10% Fetal Bovine Serum and Pennicilin / Streptomycin, at 37 degrees / 5%CO2. TalH cells (DSMZ) were used as targets for transduction to calculate viral titers. TalH cells were cultured in RP I media supplemented with 10% Fetal Bovine Serum and Pennicilin / Streptomycin, at 37 degrees / 5%CO2. CD56- cells were isolated from donor PBMCs, stimulated with EBV PepMix from JPT Peptides (Berlin, Germany), and cultured in II2, II7, and ill 5.

[0212] Lentivirus Packaging / Mutant generation

[0213] Third generation Lentivirus were produced by transfection of HEK293T or 293T-LentiX with Fugene 6 and the packaging plasmids pMDLG / pRRE (Addgene#12251), pRSV-Rev(Addgene#12253), a VSV-G plasmid, with / without an additional Affi-VSVG plasmid, and transfer plasmid containing a GFP or CAR coding sequence. VSV-Gwt was encoded by pMD2.G VSVG (Addgene#12259). Affi-VSVG were chemically synthesized by Atum (CA, USA) and cloned on to a pd633 plasmid backbone (CMV promoter). The encoded protein included the VSV-G signal sequence, 6xHistidine tag, an scFv derived from the SP34 antibody, and the remainder of the VSV-G amino acids. Mutant versions of the VSV-G and Affi-VSV-G were generated by a Site Directed Mutagenesis kit.

[0214] Viral TiteringSupernatant containing lentivirus were collected after 48 and 72h. Viral supernatants were centrifuged to pellet cellular debris and added to freshly plated TalU cells. Transduction efficiency was determined by GFP expression in live cells as detected by the Aurora 3L flow cytometer by Cytek (CA, USA). Viral titer was calculated as (% cells transduced) * (# cells plated) / volume virus added.

[0215] DETAILED DESCRIPTION OF FIGURES

[0216] Example embodiments of the disclosure will be better understood and readily apparent to one of ordinary skill in the art from the following discussions and if applicable, in conjunction with the figures. Example embodiments are not necessarily mutually exclusive as some may be combined with one or more embodiments to form new exemplary embodiments. The example embodiments should not be construed as limiting the scope of the disclosure.

[0217] FIG. 1 shows bar graphs in which the lentivirus production process was improved when VSVG1-5R replaced VSV-G. pMD2G plasmid encoding for VSV-G was mutated to produce VSVG resistant to the cellular degradation machinery (VSVGmut). The VSVG1-5R plasmid replaced VSV-G wild-type encoded by pMD2G in the lentivirus production process and produced higher titer. Virus carrying the GFP gene were packaged in 293T cells and transduced on TalU cells. TalH cells were analyzed for GFP expression by flow cytometry.

[0218] FIG. 2 shows line graphs in which Affi-VSVG increased the infectivity towards target cells. VSVG was combined with a binding domain to T cells. Peripheral Blood Monocytes containing T cells were transduced with VSVG or Affi-VSVG pseudotyped lentivirus designed to transfer the GFP gene. The percent transduction was determined by measurement of GFP expression in live cells analyzed by flow cytometry. T cell stimulation changes the level of VSV-G ligands on the cell surface but Affi-VSVG outperformed VSVG before and after stimulation.

[0219] FIG. 3 shows bar graphs in which the lentivirus production process was improved when Affi-VSVG 1-5R replaced Affi-VSVG. Lentivirus were pseudotyped with (VSVG or VSVG1-5R) and (Affi-VSVG or Affi-VSVG 1-5R), packaged in 293T cells, and titered on TalU cells which is positive for the target molecule of Affi-VSVG. Using either degradation resistant plasmid in place of its wild-type counterpart improved titer and combining the two degradation resistant envelopes produced a synergistic effect and highest titer.FIG. 4 shows a bar graph in which VSVG3.4R Affi-VSV improved virus titer. VSVG3,4R contained Lysine to Arginine mutations at residues 496 and 497, which were sufficient to confer improved titer to lentivirus pseudotyped with VSVG3.4R in place of pMD2G and tittered on Tall1 cells. The improvement was also effected when VSVG3,4R replaced pMD2G in viruses pseudotyped with Affi-VSVG or Affi-VSVG1-5R.

[0220] FIG. 5 shows a bar graph in which VSVG5R and Aff-VSVG1-5R improved virus titer. VSVG5R improved lentivirus titer compared to VSVGwt, when the viruses were pseudotyped with VSVG alone or in combination with some form of Aff-VSVG. Virus carrying the GFP gene were packaged in 293 cells and transduced on TalH cells. TalH cells were analyzed for GFP expression by flow cytometry.

[0221] FIG. 6 shows a bar graph in which VSVG1-4R and Affi-VSVG 1-5R improved virus titer. VSVG1-4R, containing Lysine to Arginine mutations at residues 491, 493, 496 and 497, improved lentivirus titer compared to VSVG, when the viruses were pseudotyped with mutant VSVG alone or in combination with some form of Affi-VSVG. Virus carrying the GFP gene were packaged in 293T cells and transduced on TalH cells. TalH cells were analyzed for GFP expression by flow cytometry.

[0222] FIG. 7 shows a bar graph in which VSVG1R (K491R) and VSVG1-2R (K491 R / K493R) do not improve lentivirus titer compared to wild-type VSV_G, when the viruses were pseudotyped with mutant VSVG alone or in combination with some form of Affi-VSVG. Virus carrying the GFP gene were packaged in 293T cells and transduced on TalH cells. TalH cells were analyzed for GFP expression by flow cytometry.

[0223] FIG. 8 shows a bar graph in which VSVG1-4R and VSVG1-5R improved virus titer for a therapeutic Chimeric Antigen Receptor (CAR) transfer vector. Virus carrying a typical CAR gene coupled to a GFP co-expression reporter gene were packaged in 293TX cells and transduced on TalH cells. TalH cells were analyzed forGFP expression by flow cytometry.

[0224] FIG. 9 shows a bar graph in which VSVG1-5N yielded similar GFP lentivirus titer as wild-type VSVG, and a bar graph in which VSVG1-5N improved lentivirus titer for a therapeutic CAR transfer vector. The Kto N mutations preserved VSV-G function in GFP carrying lentivirus and improved the virus titer for CAR carrying lentivirus when the viruses were pseudotyped with mutant VSVG alone or in combination with some form of Affi-VSVG. Viruses carrying GFP reporter gene or a typical CAR gene coupled to a GFP co-expression reporter gene were packaged in 293TX cells and transduced on TalH cells. TalH cells were analyzed for GFP expression by flow cytometry.APPLICATIONS

[0225] Embodiments of the methods disclosed herein provide a viral glycoprotein comprising a glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein and / or a modified N-terminal that allows the glycoprotein to be resistant towards detection and / or degradation.

[0226] Advantageously, the present disclosure provides a glycoprotein that when used for pseudotyping viruses generates viruses with higher titer and higher infectivity / transduction of target cell types (such as T cells).

[0227] Advantageously, the present disclosure includes transferring of a CAR gene into T cells to generate CAR-T cells / therapeutic CAR-T cells.

[0228] Even more advantageously, the present disclosure provides a glycoprotein for applications in the cell & gene therapy sector where viruses can be developed and commercialized for the treatment of gene disorders, generation of gene-modified cell therapies, in vivo injection for gene therapy and / or production of oncolytic virus for cancer treatment.

[0229] More advantageously, the present disclosure provides a glycoprotein that can be used in the pharmaceuticals / biotechnology sector / research organisations to produce viruses for therapies or processes.

[0230] It will be appreciated by a person skilled in the art that other variations and / or modifications may be made to the embodiments disclosed herein without departing from the spirit or scope of the disclosure as broadly described. For example, in the description herein, features of different exemplary embodiments may be mixed, combined, interchanged, incorporated, adopted, modified, included etc. or the like across different exemplary embodiments. The present embodiments are, therefore, to be considered in all respects to be illustrative and not restrictive.

Claims

CLAIMS1. A viral glycoprotein comprisinga glycoprotein comprising one or more mutations at the intracellular domain of the glycoprotein and / or a modified N-terminal that allows the glycoprotein to be resistant towards detection and / or degradation.

2. The glycoprotein of claim 1, wherein the one or more mutations is a substitution at the C-terminal or at positions 491 to 511.

3. The glycoprotein of any one of the preceding claims, wherein the one or more mutations is a substitution at position 491, 493, 496, 497, 511, or combinations thereof.

4. The glycoprotein of any one of the preceding claims, wherein the one or mutations is a substitution to an amino acid with a positively charged side chain and / or an amino acid with a polar uncharged side chain.

5. The glycoprotein of any one of the preceding claims, wherein the one or more mutations comprise one or more substitutions to an amino acid that is not subjected to ubiquitylation / degradation.

6. The glycoprotein of any one of the preceding claims, wherein the one or more mutations comprise one or more substitutions to an arginine (“R”) and / or to an asparagine (“N”).

7. The glycoprotein of any one of the preceding claims, wherein the modified N-terminal comprises a binding domain that binds to an immune cell.

8. The glycoprotein of any one of the preceding claims, comprising the sequence:VSVG Ctrl (pMDLG)_amino acid (SEQ ID NO: 1) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgk> VSVG1R_amino acid (SEQ ID NO: 2) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnstwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRlkh tkkrqiytdiemnrlgk> VSVG2R_amino acid (SEQ ID NO: 3) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciklRh tkkrqiytdiemnrlgk> VSVG1-2R_amino acid (SEQ ID NO: 4) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesiffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htkkrqiytdiemnrlgk> VSVG3,4R_amino acid (SEQ ID NO: 5) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht RRrqiytdiemnrlgk> VSVG1-4R_amino acid (SEQ ID NO: 6) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgk> VSVG5R_amino acid (SEQ ID NO: 7) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfsswkssiasfffiigliiglflvlrvgihlciklkht kkrqiytdiemnrlgR> VSVG1-5R_amino acid (SEQ ID NO: 8) Mkcllylaflfigvnckftivfphnqkgnwknvpsnyhycpsssdlnwhndligtalqvkmpkshkaiqadgwmchask wvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgyatvtdaeavivqvtphhvlvdeytgewv dsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelsslgkegtgfrsnyfayetggkackmqyck hwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdverildyslcqetwskiraglpispvdlsylap knpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapyedveigpngvlrtssgykfplymighg mldsdlhlsskaqvfehphiqdaasqlpddesIffgdtglsknpielvegwfsswkssiasfffiigliiglflvIrvgihlciRIR htRRrqiytdiemnrlgR,> Affi-VSVGwt / Affi-VSVG (SEQ ID NO: 19) Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsggggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvlrvgihlciklkhtkkrqiytdiemnrlgk,> Affi-VSVG1-5R (SEQ ID NO: 24) Mkcllylaflfigvnchhhhhhevqlvesggglvqpggslklscaasgftfnkyamnwvrqapgkglewvarirskynny atyyadsvkdrftisrddskntaylqmnnlktedtavyycvrhgnfgnsyisywaywgqgtlvtvssggggsggggsgg ggsqtvvtqepsltvspggtvtltcgsstgavtsgnypnwvqqkpgqaprgliggtkflapgtparfsgsllggkaaltlsgvq pedeaeyycvlwysnrwvfgggtkltvlgggsgggssgggskftivfphnqkgnwknvpsnyhycpsssdlnwhndli gtalqvkmpkshkaiqadgwmchaskwvttcdfrwygpkyithsirsftpsveqckesieqtkqgtwlnpgfppqscgy atvtdaeavivqvtphhvlvdeytgewvdsqfingkcsnyicptvhnsttwhsdykvkglcdsnlismditffsedgelssl gkegtgfrsnyfayetggkackmqyckhwgvrlpsgvwfemadkdlfaaarfpecpegssisapsqtsvdvsliqdve rildyslcqetwskiraglpispvdlsylapknpgtgpaftiingtlkyfetryirvdiaapilsrmvgmisgttterelwddwapy edveigpngvlrtssgykfplymighgmldsdlhlsskaqvfehphiqdaasqlpddeslffgdtglsknpielvegwfss wkssiasfffiigliiglflvIrvgihlciRIRhtRRrqiytdiemnrlgRor an amino acid sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above, or an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid differences thereof.

9. The glycoprotein of any one of the preceding claims, encoded by a sequence comprising> VSVG Ctrl (pMDLG)_nt (SEQ ID NO: 9) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatagagatgaaccgacttggaaag> VSVG1R_nt (SEQ ID NO: 10) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtgaagca caccaagaaaagacagatttatacagacatagagatgaaccgacttggaaag> VSVG2R_nt (SEQ ID NO: 11)atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattGaGAcaca cAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag> VSVG1-2R_nt (SEQ ID NO: 12) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcatCCGGCtGaGAc acacAaagaaaagacagatttatacagacatagagatgaaccgacttggaaag> VSVG3,4R_nt (SEQ ID NO: 13) atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caGACGCagacagatttatacagacatagagatgaaccgacttggaaag> VSVG1-4R_nt (SEQ ID NO: 14) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatagagatgaaccgacttggaaag> VSVG5R_nt (SEQ ID NO: 15) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaagcacac caagaaaagacagatttatacagacatCgagatgaaccgGctGggCaGA> VSVG1-5R_nt (SEQ ID NO: 16)atgaagtgccttttgtacttagcctttttattcattggggtgaattgcaagttcaccatagtttttccacacaaccaaaaaggaa actggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacagcct tacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcactactt gtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaag cattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacggatg ccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacagttcat caacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaagggc tatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcac agggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattggggagtc agactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaagggt caagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctctgcc aagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaacccagga accggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgctccaat cctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatatgaaga cgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggtatgttgg actccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctgatgatg agagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaagctctat tgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtGaGAca cacAaGACGCagacagatttatacagacatCgagatgaaccgGctGggCaGA> Affi-VSVGwt_nt (SEQ ID NO: 17) Atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgattcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaaggaaagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagg gcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgttggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattaaattaaa gcacaccaagaaaagacagatttatacagacatagagatgaaccgacttggaaagtaa> Affi-VSVG1-5R_nt (SEQ ID NO: 18) atgaagtgccttttgtacttagcctttttattcattggggtgaattgccatcaccatcaccatcacgaggtgcagctggtcgagt ctggaggaggattggtgcagcctggagggtcattgaaactctcatgtgcagcctctggattcaccttcaataagtacgcca tgaactgggtccgccaggctccaggaaagggtttggaatgggttgctcgcataagaagtaaatataataattatgcaaca tattatgccgatcagtgaaagacaggttcaccatctccagagatgattcaaaaaacactgcctatctacaaatgaacaa cttgaaaactgaggacactgccgtgtactactgtgtgagacatgggaacttcggtaatagctacatatcctactgggcttac tggggccaagggactctggtcaccgtctcctcaggtggtggtggttctggcggcggcggctccggtggtggtggttctcag actgttgtgactcaggaaccttcactcaccgtatcacctggtggaacagtcacactcacttgtggctcctcgactggggctgt tacatctggcaactacccaaactgggtccaacaaaaaccaggtcaggcaccccgtggtctaataggtgggactaagttc ctcgcccccggtactcctgccagattctcaggctccctgcttggaggcaaggctgccctcaccctctcaggggtacagcc agaggatgaggcagaatattactgtgttctatggtacagcaaccgctgggtgttcggtggaggaaccaaactgactgtcct aggcggcggcagcggcggcggcagcagcggcggcggcagcaagttcaccatagtttttccacacaaccaaaaagg aaactggaaaaatgttccttctaattaccattattgcccgtcaagctcagatttaaattggcataatgacttaataggcacag ccttacaagtcaaaatgcccaagagtcacaaggctattcaagcagacggttggatgtgtcatgcttccaaatgggtcact acttgtgatttccgctggtatggaccgaagtatataacacattccatccgatccttcactccatctgtagaacaatgcaagga aagcattgaacaaacgaaacaaggaacttggctgaatccaggcttccctcctcaaagttgtggatatgcaactgtgacg gatgccgaagcagtgattgtccaggtgactcctcaccatgtgctggttgatgaatacacaggagaatgggttgattcacag ttcatcaacggaaaatgcagcaattacatatgccccactgtccataactctacaacctggcattctgactataaggtcaaa gggctatgtgattctaacctcatttccatggacatcaccttcttctcagaggacggagagctatcatccctgggaaaggagggcacagggttcagaagtaactactttgcttatgaaactggaggcaaggcctgcaaaatgcaatactgcaagcattgggg agtcagactcccatcaggtgtctggttcgagatggctgataaggatctctttgctgcagccagattccctgaatgcccagaa gggtcaagtatctctgctccatctcagacctcagtggatgtaagtctaattcaggacgttgagaggatcttggattattccctc tgccaagaaacctggagcaaaatcagagcgggtcttccaatctctccagtggatctcagctatcttgctcctaaaaaccc aggaaccggtcctgctttcaccataatcaatggtaccctaaaatactttgagaccagatacatcagagtcgatattgctgct ccaatcctctcaagaatggtcggaatgatcagtggaactaccacagaaagggaactgtgggatgactgggcaccatat gaagacgtggaaattggacccaatggagttctgaggaccagttcaggatataagtttcctttatacatgattggacatggta tgtggactccgatcttcatcttagctcaaaggctcaggtgttcgaacatcctcacattcaagacgctgcttcgcaacttcctg atgatgagagtttattttttggtgatactgggctatccaaaaatccaatcgagcttgtagaaggttggttcagtagttggaaaa gctctattgcctcttttttctttatcatagggttaatcattggactattcttggttctccgagttggtatccatctttgcattCGGCtG aGAcacacAaGACGCagacagatttatacagacatagagatgaaccgGctGggCaGA,or polynucleotide sequence having at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.75%, or 100% sequence identity to any one of the sequences above.

10. The glycoprotein of any one of the preceding claims comprising a combination of glycoproteins selected from any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17 (Affi-VSVGwt), SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); andany one of the glycoproteins with sequences: SEQ ID NO: 2 (VSVG1R), SEQ ID NO: 3 (VSVG2R), SEQ ID NO: 4 (VSVG1-2R), SEQ ID NO: 5 (VSVG3.4R), SEQ ID NO: 6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R) or SEQ ID NO: 1 (VSVG (wild type / control)).

11. The glycoprotein of any one of the preceding claims comprising a combination of glycoproteins selected from any one of the glycoproteins with nucleotide sequences SEQ ID NO: 17 (Affi-VSVGwt) or SEQ ID NO: 18 (Affi-VSVG1-5R) or amino acid sequences SEQ ID NO: 19 (AffiVSVGwt), SEQ ID NO: 24 (Affi-VSVG1-5R); andany one of the glycoproteins with sequences: SEQ ID NO: 5 (VSVG3,4R), SEQ ID NO: 6 (VSVG1-4R), SEQ ID NO: 7 (VSVG5R), SEQ ID NO: 8 (VSVG1-5R), or SEQ ID NO: 1 (VSVG (wild type / control)).

12. The glycoprotein of any one of the preceding claims comprising the combination of glycoproteins with sequences: SEQ ID NO: 8 (VSVG1-5R) and SEQ ID NO: 18 (Affi-VSVG1-5R).

13. A polynucleotide encoding any one of the glycoproteins of any one of the preceding claims.

14. A vector encoding the polynucleotide or glycoprotein of any one of the preceding claims.

15. A host cell comprising the polynucleotide, the vector, or glycoprotein of any one of the preceding claims.

16. A composition comprising the glycoprotein or polynucleotide of any one of the preceding claims.

17. The glycoprotein of any one of the preceding claims for use in a therapy, optionally wherein the therapy comprises gene therapy.

18. A method of treating a disease, comprising providing the glycoprotein of any one of the preceding claims.

19. Use of the glycoprotein of any one of the preceding claims in the manufacture of a medicament for treating a disease, optionally wherein the medicament is a gene therapy.

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