A gE protein antigen variant, its preparation method and its application

Abstract

This invention discloses a gE protein antigen variant, its preparation method, and its applications. Specifically, the invention provides a gE protein antigen variant comprising a truncated C-terminus of any amino acid residue from amino acid residues 547 to 623 of the gE protein. The invention also provides a nucleic acid molecule encoding the aforementioned gE protein antigen variant, a vector containing the aforementioned nucleic acid molecule, a cell containing the aforementioned vector, and a pharmaceutical composition containing the aforementioned nucleic acid molecule, vector, and cell. Furthermore, this invention provides a vaccine and a derivative of the gE protein antigen variant. This invention provides the application of the aforementioned gE protein antigen variant in the preparation of products for detecting varicella-zoster virus and in the preparation, capture, or screening of antibodies that recognize the varicella-zoster virus gE protein. The gE protein antigen variant disclosed in this invention has high application value and broad market prospects.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and relates to a gE protein antigen variant, its preparation method, and its application. Background Technology

[0002] Herpes zoster (HZ) is an acute infectious skin disease caused by the varicella-zoster virus (VZV). After infecting the body, the VZV virus can remain latent in the neurons of the dorsal root ganglia of the spinal cord for a long time. When the body's immune function is weakened, VZV can be reactivated and spread along nerve fibers to the skin, causing a strong inflammatory response in the infected nerve and skin systems, thus triggering herpes zoster. HZ usually manifests as a painful and itchy rash on one side of the body, lasting for two to four weeks. The risk of developing the disease increases with age. A common complication is postherpetic neuralgia, where the pain can last from months to years, severely affecting the patient's normal work and life.

[0003] Vaccination against shingles is an important means of reducing the incidence of shingles. Globally available shingles vaccines are mainly live attenuated shingles vaccines (ZVL) and recombinant shingles vaccines (RZV).

[0004] One is a live attenuated vaccine developed by Merck, approved in 2006, marketed under the brand name Zostervax; the other is a live attenuated vaccine developed by Changchun BCHT, approved in 2023, marketed under the brand name HZV-live. The third is a subunit adjuvanted vaccine developed by GlaxoSmithKline (GSK), marketed under the brand name Shingrix, which was first approved in North America in 2017. However, both vaccines have shortcomings in efficacy.

[0005] The live attenuated vaccine uses the varicella-zoster virus Oka strain, the same strain used in the varicella vaccine, but with a potency more than 10 times higher. A drawback of this vaccine is that it can establish a latent infection in the ganglia, which can then reactivate and cause shingles. Furthermore, live attenuated viruses are not suitable for all subjects, including immunocompromised patients, such as those with leukemia, lymphoma, systemic malignancies, immunodeficiency diseases, or those undergoing immunosuppressive therapy—groups at high risk for shingles. Merck's Zostavax live attenuated vaccine reduced the incidence of shingles by 69.8% in immunocompetent individuals aged 50–59 years, while in those ≥60 years, it reduced the incidence of shingles, PHN, and disease burden by 51.3%, 66.5%, and 61.1%, respectively. The preventive efficacy of Zostavax gradually decreases with age, and severe immunosuppression and pregnancy are contraindications. Therefore, the need to develop safer and more effective vaccines is particularly urgent.

[0006] GSK's VZV gE recombinant protein and AS01B adjuvant-based shingles subunit vaccine reduced the incidence of shingles and herpes zoster (PHN) by 97.2% and 91.2% respectively in immunocompetent individuals aged ≥50 years, and by 89.8% and 88.8% respectively in individuals aged ≥70 years, demonstrating superior efficacy compared to live attenuated vaccines. A drawback of Shingrix is ​​its inherent inadequacy as a subunit vaccine in activating cellular immune responses, requiring the use of an adjuvant. Clinical studies have found that the protective effect against shingles is primarily related to vaccine-induced CD4+ activation. + T-cell immune responses are involved. However, subunit vaccines are weaker in activating cellular immune responses. To induce a stronger cellular immune response than live attenuated vaccines, highly potent adjuvants, such as AS01B, must be added. The addition of AS01B increases the toxicity of the Shingrix vaccine. It has been reported that in Shingrix clinical studies, more than 10% of subjects experienced grade 3 adverse reactions, including headaches and pain. Furthermore, the key component of AS01B, QS21, is a polysaccharide mixture that can only be extracted from the bark of the soapberry tree, which grows in temperate regions of South America, using reversed-phase high-performance liquid chromatography. It cannot currently be synthesized artificially, resulting in limitations such as limited sourcing, difficulties in quality control during preparation, and the need for detoxification agents due to its hemolytic activity. This leads to the high price of Shingrix (US$150-200 per dose, requiring two doses) and a supply shortage. Summary of the Invention

[0007] To address the technical problems existing in the prior art, the following technical solutions are provided:

[0008] This invention provides a gE protein antigen variant comprising a truncated C-terminus of any amino acid residue selected from amino acid residues at positions 547 to 623 of the gE antigen, wherein the position of the amino acid residue is referenced to the amino acid sequence SEQ ID NO:1 (MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGES).

[0009] In embodiments of the present invention, the truncation of the carboxyl terminus of the Xth amino acid residue refers to retaining the first to the Xth amino acid residues in the direction from the amino terminus (N-terminus) to the carboxyl terminus (C-terminus), and truncating the continuous amino acid residues from the (X+1)th amino acid residue to the carboxyl terminus. In a specific embodiment of the present invention, the "amino acid at position "X" refers to the amino acid corresponding to the amino acid at position "X" in the gE protein of the VZV oka strain of SEQ ID NO: 1. Note that when using a VZV strain instead of an oka strain, the amino acid positions of the gE protein will be numbered by inserting gaps where necessary to align the sequence of other RSV strains with the gE protein of SEQ ID NO: 1, referring to the numbering of the gE protein of the oka strain of SEQ ID NO: 1. Sequence alignment can be performed using methods well known in the art, such as CLUSTALW, Bioedit, or CLC Workbench.

[0010] In some specific embodiments, the gE protein antigen variant is selected from positions 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 5... 85th, 586th, 587th, 588th, 589th, 590th, 591st, 592nd, 593rd, 594th, 595th, 596th, 597th, 598th, 599th, 600th, 601st, 602nd, 603rd, 604th, 605th, 60th The carboxyl terminus of any amino acid residue at positions 6, 607, 608, 609, 610, 611, 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622, and 623 is truncated.

[0011] The term "amino acid residue" specifically includes alanine (Ala or A), cysteine ​​(Cys or C), aspartic acid (Asp or D), glutamic acid (Glu or E), phenylalanine (Phe or F), glycine (Gly or G), histidine (His or H), isoleucine (Ile or I), lysine (Lys or K), leucine (Leu or L), methionine (Met or M), asparagine (Asn or N), proline (Pro or P), glutamine (Gln or Q), arginine (Arg or R), serine (Ser or S), threonine (Thr or T), valine (Val or V), tryptophan (Trp or W), and tyrosine (Tyr or Y) residues. The term "amino acid residue" may also include non-standard, unconventional, or non-natural amino acids, and may optionally include amino acids other than any of the following: alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, and tyrosine residues. The term "amino acid residue" may also include α-, β-, γ-, and δ-amino acids.

[0012] Furthermore, the gE protein antigen variant includes a polypeptide truncated at the carboxyl terminus of amino acid residues selected from positions 547, 551, 556, 561, 566, 571, 576, 581, 586, 591, 596, 601, 606, 611, 616, and 621.

[0013] Furthermore, the amino acid sequence of the gE protein antigen variant has at least 90%, preferably at least 95%, preferably at least 96%, preferably at least 97%, preferably at least 97%, preferably at least 98%, and preferably at least 99% identity with the amino acid sequence shown in any of SEQ ID NO:2-18.

[0014] Furthermore, the gE protein antigen variant is selected from the following amino acid sequence:

[0015] SEQ ID NO:2 (Truncation of the carboxyl terminus of amino acid residue 547 of gE antigen):

[0016] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0017] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0018] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0019] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0020] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0021] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0022] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAA;

[0023] SEQ ID NO:3 (Truncation at the carboxyl terminus of amino acid residue 551 of the gE antigen):

[0024] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0025] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0026] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0027] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0028] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0029] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0030] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLL;

[0031] SEQ ID NO:4 (Truncation at the carboxyl terminus of amino acid residue 556 of gE antigen):

[0032] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0033] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0034] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0035] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0036] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0037] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0038] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIF;

[0039] SEQ ID NO:5 (Truncation at the carboxyl terminus of amino acid residue 561 of the gE antigen):

[0040] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0041] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0042] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0043] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0044] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0045] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0046] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTA;

[0047] SEQ ID NO:6 (Truncation at the carboxyl terminus of amino acid residue 566 of the gE antigen):

[0048] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0049] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0050] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0051] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0052] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0053] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0054] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRV;

[0055] SEQ ID NO:7 (Truncation of the carboxyl terminus of amino acid residue 571 of gE antigen):

[0056] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0057] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0058] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0059] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0060] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0061] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0062] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRV;

[0063] SEQ ID NO:8 (Truncation of the carboxyl terminus of amino acid residue 576 of gE antigen)

[0064] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0065] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0066] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0067] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0068] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0069] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0070] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIEERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPY;

[0071] SEQ ID NO:9 (C-terminal truncation of the 581st amino acid residue of the gE antigen):

[0072] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0073] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0074] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0075] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0076] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0077] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0078] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0079] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMY;

[0080] SEQ ID NO:10(truncation at the carboxyl terminus of amino acid residue 586 of the gE antigen):

[0081] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0082] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0083] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0084] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0085] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0086] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0087] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0088] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLP;

[0089] SEQ ID NO:11(truncation at the carboxyl terminus of amino acid residue 591 of gE antigen):

[0090] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0091] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0092] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0093] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0094] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0095] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0096] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0097] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFE;

[0098] SEQ ID NO:12 (truncation at the carboxyl terminus of amino acid residue 596 of the gE antigen):

[0099] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0100] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0101] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0102] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0103] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0104] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0105] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0106] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSEST;

[0107] SEQ ID NO:13(truncation at the carboxyl terminus of amino acid residue 601 of gE antigen):

[0108] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0109] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0110] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0111] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0112] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0113] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0114] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0115] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEE);

[0116] SEQ ID NO:14(truncation at the carboxyl terminus of amino acid residue 606 of the gE antigen):

[0117] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0118] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0119] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0120] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0121] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0122] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0123] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0124] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAI;

[0125] SEQ ID NO:15 (Truncation of the carboxyl terminus of amino acid residue 611 of gE antigen):

[0126] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0127] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0128] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0129] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0130] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0131] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0132] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0133] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHG;

[0134] SEQ ID NO:16 (Truncation of the carboxyl terminus of amino acid residue 616 of gE antigen):

[0135] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0136] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0137] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0138] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0139] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0140] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0141] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0142] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYT;

[0143] SEQ ID NO:17(truncation at the carboxyl terminus of amino acid residue 621 of the gE antigen):

[0144] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0145] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR

[0146] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0147] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0148] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0149] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0150] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0151] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDK;

[0152] SEQ ID NO:18 (C-terminal truncation of the 623rd amino acid residue of the gE antigen)

[0153] MGTVNKPVVGVLMGFGIITGTLRITNPVRASVLRYDDFHIDEDKLDTNSVYEPYYHSDHAESSWVNRGESSR

[0154] KAYDHNSPYIWPRNDYDGFLENAHEHHGVYNQGRGIDSGERLMQPTQMSAQEDLGDDTGIHVIPTLNGDDR [[ID=二十九]]

[0155] HKIVNVDQRQYGDVFKGDLNPKPQGQRLIEVSVEENHPFTLRAPIQRIYGVRYTETWSFLPSLTCTGDAAPAI

[0156] QHICLKHTTCFQDVVVDVDCAENTKEDQLAEISYRFQGKKEADQPWIVVNTSTLFDELELDPPEIEPGVLKVL

[0157] RTEKQYLGVYIWNMRGSDGTSTYATFLVTWKGDEKTRNPTPAVTPQPRGAEFHMWNYHSHVFSVGDTFSLA

[0158] MHLQYKIHEAPFDLLLEWLYVPIDPTCQPMRLYSTCLYHPNAPQCLSHMNSGCTFTSPHLAQRVASTVYQNC

[0159] EHADNYTAYCLGISHMEPSFGLILHDGGTTLKFVDTPESLSGLYVFVVYFNGHVEAVAYTVVSTVDHFVNAIE

[0160] ERGFPPTAGQPPATTKPKEITPVNPGTSPLLRYAAWTGGLAAVVLLCLVIFLICTAKRMRVKAYRVDKSPYNQSMYYAGLPVDDFEDSESTDTEEEFGNAIGGSHGGSSYTVYIDKTR;

[0161] Furthermore, the gE protein antigen variant is selected from a truncated C-terminus of the 571st amino acid residue, and the amino acid sequence is shown in SEQ ID NO:7.

[0162] Furthermore, the gE protein antigen variant is selected from a truncated C-terminus of the 591st amino acid residue, and the amino acid sequence is shown in SEQ ID NO:11.

[0163] As used herein, the term "antigen variant" refers to a polypeptide sequence that differs from the specifically recognized sequence, wherein one or more amino acid residues are deleted, substituted, or added. Variants can be naturally occurring allelic antigen variants or non-natural antigen variants. Variants may originate from the same species or from other species and may include homologs, paralogs, and orthologs. In some embodiments, antigen variants of polypeptides useful in this invention have the same or similar biological activities as those of the parent polypeptide, including hormonal function or antigen-binding properties. The term "antigen variant" with respect to (poly)peptides includes all forms of polypeptides as defined herein. The term "antigen variant" includes naturally occurring polypeptides as well as recombinant and synthetically produced polypeptides.

[0164] The present invention provides a nucleic acid molecule comprising a nucleotide sequence encoding the aforementioned gE protein antigen variant.

[0165] The terms “nucleic acid molecule,” “coding sequence,” “polynucleotide,” “nucleotide sequence,” “nucleic acid sequence,” and “nucleic acid” are used interchangeably and include DNA, RNA, or their hybrids, which can be double-stranded or single-stranded.

[0166] The present invention provides a carrier comprising the nucleic acid molecules described above.

[0167] Furthermore, the vector includes plasmid vectors, lentiviral vectors, adenovirus vectors, adeno-associated virus vectors, piggyBac vectors, or Sleeping Beauty transposable vectors.

[0168] The term "vector" is used to refer to a nucleic acid molecule that can be inserted to introduce a nucleic acid sequence into a cell, where it can replicate. The nucleic acid sequence can be "exogenous," meaning it is foreign to the cell into which the vector is introduced, or the sequence is homologous to a sequence already present in the cell. Vectors include plasmids, granules, viruses (bacteriophages, animal viruses, and plant viruses), and artificial chromosomes (e.g., YAC).

[0169] The present invention provides a cell comprising the aforementioned gE protein antigen variant, the aforementioned nucleic acid molecule, and the aforementioned vector.

[0170] Furthermore, the cells include prokaryotic cells or eukaryotic cells.

[0171] Furthermore, the prokaryotic cells include bacteria and actinomycetes.

[0172] Furthermore, the eukaryotic cells include mammalian cells, insect cells, and yeast cells.

[0173] In some specific embodiments, the prokaryotic cells include bacteria, such as *Escherichia coli*, *Bacillus subtilis*, *Salmonella*, *Pseudomonas*, *Streptomyces*, and *Staphylococcus*. In some specific embodiments, the mammalian cells include fibroblasts, lymphocytes, epithelial cells, and myeloid cells.

[0174] Furthermore, the eukaryotic cells include 293T cells.

[0175] The term "cell" refers to a prokaryotic or eukaryotic cell and its progeny, that is, any and all progeny. It includes any transformable organism capable of replicating a vector and / or expressing a heterologous gene encoded by the vector. Cells can and have been used as recipients of vectors. Cells can be "transfected" or "transformed," which refers to the process of transferring or introducing exogenous nucleic acids into a cell; transformed cells include primary cells and their progeny.

[0176] The present invention provides a pharmaceutical composition comprising the aforementioned gE protein antigen variant, the aforementioned nucleic acid molecule, the aforementioned carrier, or the aforementioned cell.

[0177] The present invention provides a vaccine comprising an active ingredient comprising an RNA polynucleotide having an open reading frame encoding at least one of the aforementioned gE protein antigen variants, and / or the aforementioned gE protein antigen variants.

[0178] Furthermore, the vaccine includes RNA vaccines, protein vaccines, or viral vector vaccines.

[0179] In a preferred embodiment, the vaccine is an RNA vaccine comprising an RNA polynucleotide having an open reading frame encoding at least the aforementioned gE protein antigen variant.

[0180] Furthermore, the open reading frames of the aforementioned RNA polynucleotides are codon-optimized. Codon optimization methods are known in the art and can be used as provided herein. In some embodiments, codon optimization can be used to match codon frequencies in the target and host organisms to ensure proper folding; to bias GC content to increase mRNA stability or reduce secondary structures; to minimize the execution of tandem repeat codons or bases that could impair gene structure or expression; to customize transcription and translation control regions; to insert or remove protein transport sequences; to remove / add post-translational modification sites (e.g., glycosylation sites) in encoded proteins; to add, remove, or replace protein domains; to insert or delete restriction sites; to modify ribosome binding sites and mRNA degradation sites; to modulate translation rates so that various protein domains can fold properly; or to reduce or eliminate problematic secondary structures within polynucleotides.

[0181] In some embodiments, codon-optimized RNA can be RNA in which the G / C level is enhanced. The G / C content of a nucleic acid molecule (e.g., mRNA) can affect RNA stability. RNA with increased amounts of guanine (G) and / or cytosine (C) residues can be functionally more stable than RNA containing large amounts of adenine (A) and thymine (T) or uracil (U) nucleotides. For example, WO02 / 098443 discloses a pharmaceutical composition containing mRNA stabilized in the translation region by sequence modification. Due to genetic code degradation, modification works by replacing existing codons with those that promote greater RNA stability without altering the resulting amino acids. The method is limited to the coding region of RNA.

[0182] Furthermore, the RNA polynucleotide contains at least one chemical modification.

[0183] Furthermore, the chemical modification is selected from any group of the following: pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methylpseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methylpseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2′-O-methyluridine.

[0184] Furthermore, at least 50% of the uracil in the RNA polynucleotide is chemically modified.

[0185] Furthermore, at least 60% of the uracil in the RNA polynucleotide is chemically modified.

[0186] Furthermore, at least 70% of the uracil in the RNA polynucleotide is chemically modified.

[0187] Furthermore, at least 80% of the uracil in the RNA polynucleotide is chemically modified.

[0188] Furthermore, at least 90% of the uracil in the RNA polynucleotide is chemically modified.

[0189] Furthermore, at least 100% of the uracil in the RNA polynucleotide is chemically modified.

[0190] Furthermore, the chemical modification is located at the 5th position of uracil.

[0191] Furthermore, the RNA polynucleotide also includes a 3'-UTR and / or at least one 5'-UTR.

[0192] Furthermore, the RNA polynucleotide also includes a 3'-UTR and at least one 5'-UTR.

[0193] Furthermore, each of the at least 3'-UTR and the at least 5'-UTR is heterogeneous to the other.

[0194] Furthermore, the at least one 3'UTR is derived from a gene selected from the group consisting of: housekeeping genes, genes encoding membrane proteins, genes involved in cell metabolism, genes involved in transcription, translation and replication, genes involved in protein modification and genes involved in cell division.

[0195] Furthermore, at least one 5'-UTR is derived from a gene selected from the group consisting of: housekeeping genes, genes encoding membrane proteins, genes involved in cell metabolism, genes involved in transcription, translation and replication, genes involved in protein modification and genes involved in cell division.

[0196] Furthermore, the RNA polynucleotide is capped.

[0197] Furthermore, the capping includes cap0, cap1, and cap2.

[0198] Furthermore, the capping is performed via enzymatic capping or co-transcriptional capping.

[0199] Furthermore, the enzymatic capping method involves capping with a capping enzyme, GTP, and S-adenosylmethionine (SAM).

[0200] Furthermore, the co-transcriptional capping includes the addition of a cap-like structure.

[0201] Furthermore, the cap structure analogues include 7mG(5′)ppp(5′)N1mpNp or 7mG(5′)ppp(5′)N1mpNp.

[0202] Furthermore, the RNA polynucleotide is subjected to tailing treatment.

[0203] Furthermore, the tailing includes tailing via cleavage and polyadenylation of specific factors, cleavage stimulating factors, cleavage factor I and cleavage factor II, poly(A) polymerase, poly(A) binding protein, and paired protein.

[0204] Furthermore, the added tail includes a polyadenylated tail.

[0205] Furthermore, the RNA polynucleotide is subjected to capping and tailing treatment.

[0206] Furthermore, the RNA polynucleotide has a 5′ cap, an open reading frame encoding at least one of the aforementioned gE protein antigen variants, and a 3′ polyadenylate tail.

[0207] Furthermore, the 5′ end cap contains 7mG(5′)ppp(5′)N1mpNp or contains 7mG(5′)ppp(5′)N1mpNp.

[0208] Furthermore, the polyadenylated tail has a length of about 20 to about 300 adenine nucleotides, preferably about 40 to about 200 adenine nucleotides, more preferably about 50 to about 100 adenine nucleotides, and even more preferably about 60 to about 70 adenine nucleotides.

[0209] Embodiments of the present invention provide RNA (e.g., mRNA) vaccines comprising at least one polynucleotide encoding at least one VZV antigenic polypeptide. The VZV RNA vaccines provided herein can be used to elicit a balanced immune response, encompassing both cellular and humoral immunity, without the various risks associated with DNA vaccines and live attenuated vaccines. Various RNA (e.g., mRNA) vaccines disclosed herein have generated immune responses in BALB / c mice, the results of which are discussed in detail in the Examples section. Specifically, RNA (e.g., mRNA) polynucleotide vaccines having open reading frames encoding one or more of various VZV antigens elicit a significant immune response relative to conventional VZV vaccines (e.g., attenuated VZV virus).

[0210] Furthermore, the RNA vaccine is formulated and delivered on a payload.

[0211] Furthermore, the load includes dendritic cells, cationic nanoemulsion (CNE), cationic peptides and polymers, lipopolyplex (LPR), lipopolyplex (LPR), and lipid nanoparticles (LNP).

[0212] Furthermore, the support is selected from lipid nanoparticles (LNPs).

[0213] Furthermore, the LNP includes cationic lipids, PEG-modified lipids, sterols, and non-cationic lipids.

[0214] Furthermore, the cationic lipid is selected from the group consisting of: 2,2-dilinole-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinole-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and 9-((4-(dimethylamino)butyryl)oxy)heptadecanedioic acid di((Z)-non-2-en-1-yl) ester (L319).

[0215] In certain specific embodiments, the lipid nanoparticle (LNP) formulations significantly enhance the efficacy of mRNA vaccines, including both chemically modified and unmodified mRNA vaccines. The efficacy of mRNA vaccines formulated in LNPs was tested in vivo using several different antigens. The results presented herein demonstrate the unexpected superior efficacy of mRNA vaccines formulated in LNPs compared to other vaccines. In some embodiments, the LNPs may be hydrophilic polymer particles; in other embodiments, the LNPs may be hydrophobic polymer particles. In some embodiments, the LNPs may be biodegradable cationic lipids of rapidly eliminating lipid nanoparticles (reLNPs) used to replace cationic lipids in improving the LNP formulation.

[0216] In some embodiments, lipid nanoparticles can be engineered to modify their surface properties, thereby enabling them to permeate mucosal barriers. The engineered lipid nanoparticles for mucus permeation may comprise polymeric materials (e.g., a polymeric core) and / or polymer-vitamin conjugates and / or triblock copolymers. Polymeric materials may include (but are not limited to) polyamines, polyethers, polyamides, polyesters, polyurethanes, polyureas, polycarbonates, polystyrene, polyimides, polysulfones, polyurethanes, polyacetylene, polyethylene, polyethyleneimine, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitrile, and polyarylates. The polymeric materials may be biodegradable and / or biocompatible.

[0217] In embodiments of the invention, the terms "chemically modified" and "chemically modified" refer to modifications of at least one of adenosine (A), guanosine (G), uridine (U), thymidine (T), or cytosine (C) ribonucleoside or deoxyribonucleoside in terms of its position, pattern, percentage, or population. Typically, these terms do not refer to modifications of the ribonucleotides of the cap portion of naturally occurring 5′-terminal mRNA.

[0218] Modification of polynucleotides includes (but is not limited to) those modifications described herein and includes (but is explicitly not limited to) those involving chemical modification. Polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) may contain naturally occurring, non-naturally occurring modifications, or a combination of naturally occurring and non-naturally occurring modifications. Polynucleotides may include, for example, any suitable modification to sugars, nucleobases, or nucleoside linkages (e.g., to linked phosphate esters, phosphodiester bonds, or the phosphodiester backbone).

[0219] Modified nucleotide base pairings encompass not only standard adenosine-thymine, adenosine-uracil, or guanosine-cytosine base pairs, but also base pairs formed between nucleotides and / or modified nucleotides, including non-standard or modified bases, wherein the arrangement of hydrogen bond donors and acceptors allows hydrogen bonding between non-standard and standard bases or between two complementary non-standard base structures, such as in polynucleotides having at least one chemical modification. An example of such non-standard base pairings is the modified nucleotide inosine paired with adenine, cytosine, or uracil. Any combination of base / sugar or linker can be incorporated into the polynucleotides of this invention.

[0220] In some embodiments of the present invention, the RNA vaccine comprises at least one ribonucleic acid (RNA) polynucleotide having an open reading frame encoding at least one VZV antigenic polypeptide or an immunogenic fragment thereof, wherein at least 50% (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, 100%) of the uracil in the open reading frame is chemically modified, optionally wherein the vaccine is formulated in lipid nanoparticles. In some embodiments, 100% of the uracil in the open reading frame is chemically modified. In some embodiments, the chemical modification is located at the 5th position of the uracil.

[0221] The modification (including, but not limited to, chemical modification) of polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) in the compositions, vaccines, methods, and synthetic methods applicable to the present invention includes, but is not limited to, the following or combinations thereof: 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine; 2-methylthio-N6-methyladenosine; 2-methylthio-N6-threonylcarbamoyladenosine; N6-glycinylcarbamoyladenosine; N6-isopentenyladenosine; N6-methyladenosine; N6-threonylcarbamoyladenosine; 1,2′-O-dimethyladenosine; 1-methyladenosine; 2′-O-methyladenosine; 2′-O-ribosyladenosine (phosphate ester); 2-methyladenosine; 2-methylthio-N6 Isopentenyl adenosine; 2-methylthio-N6-hydroxyn-valinecarbamoyl adenosine; 2′-O-methyl adenosine; 2′-O-ribosyl adenosine (phosphate ester); isopentenyl adenosine; N6-(cis-hydroxyisopentenyl) adenosine; N6,2′-O-dimethyl adenosine; N6,2′-O-dimethyl adenosine; N6,N6,2′-O-trimethyl adenosine; N6,N6-dimethyl adenosine; N6-acetyl adenosine; N6-hydroxyn-valinecarbamoyl adenosine; N6-methyl-N6-threonylcarbamoyl adenosine; 2-methyl adenosine; 2-methylthio-N6-isopentenyl adenosine; 7-deaza-adenosine; N1-methyl adenosine; N6,N6(dimethyl) adenosine; N6-cis-hydroxyisopentenyl adenosine Alkenyl-adenosine; α-Thio-adenosine; 2(amino)adenosine; 2(aminopropyl)adenosine; 2(methylthio)N6(isopentenyl)adenosine; 2-(alkyl)adenosine; 2-(aminoalkyl)adenosine; 2-(aminopropyl)adenosine; 2-(halo)adenosine; 2-(halo)adenosine; 2-(propyl)adenosine; 2′-amino-2′-deoxy-ATP; 2′-azido-2′-deoxy-ATP; 2′-deoxy-2′-a-aminoadenosine TP; 2′-deoxy-2′-a-azido-adenosine TP; 6(alkyl)adenosine; 6(methyl)adenosine; 6-(alkyl)adenosine; 6-(methyl)adenosine; 7(deaza)adenosine; 8(alkenyl)adenosine; 8(alkynyl)adenosine 8-(Alkyl)adenine; 8-(amino)adenine; 8-(thioalkyl)adenine; 8-(alkenyl)adenine; 8-(alkyl)adenine; 8-(alkynyl)adenine; 8-(amino)adenine; 8-(haloyl)adenine; 8-(hydroxy)adenine; 8-(thioalkyl)adenine; 8-(thiol)adenine; 8-azido-adenine; azaadenine; deazaadenine; N6(methyl)adenine; N6-(isopentyl)adenine; 7-deaza-8-azaadenine; 7-methyladenine; 1-deazaadenine TP; 2′-fluoro-N6-Bz-deoxyadenine TP; 2′-OMe-2-amino-ATP; 2′-O-methyl-N6-Bz-deoxyadenine TP; 2′-α-ethynyladenine TP;2-Aminoadenine; 2-Aminoadenosine TP; 2-Amino-ATP; 2′-α-trifluoromethyladenosine TP; 2-azidoadenosine TP; 2′-β-ethynyladenosine TP; 2-bromoadenosine TP; 2′-β-trifluoromethyladenosine TP; 2-chloroadenosine TP; 2′-deoxy-2′,2′-difluoroadenosine TP; 2′-deoxy-2′-α-mercaptoadenosine TP; 2′-deoxy-2′-α-thiomethoxyadenosine TP; 2′-deoxy-2′-β-aminoadenosine TP; 2′-deoxy-2′-β-azidoadenosine TP; 2′-deoxy-2′-β-bromoadenosine TP; 2′-deoxy-2′-β-chloroadenosine TP; 2′- Deoxy-2′-β-iodoadenosine TP; 2′-deoxy-2′-β-mercaptoadenosine TP; 2′-deoxy-2′-β-thiomethoxyadenosine TP; 2-fluoroadenosine TP; 2-iodoadenosine TP; 2-mercaptoadenosine TP; 2-methoxy-adenosine; 2-methylthio-adenosine; 2-trifluoromethyladenosine TP; 3-deaza-3-bromoadenosine TP; 3-deaza-3-chloroadenosine TP; 3-deaza-3-fluoroadenosine TP; 3-deaza-3-iodoadenosine TP; 3-deazaadenosine TP; 4′-azidoadenosine TP; 4′-carbocyclic adenosine TP; 4′-ethynyladenosine TP; 5′-homo-adenosine TP; 8-aza-ATP; 8-bromo-adenosine TP; 8-trifluoromethyladenosine TP; 9-Dezaaadenosine (TP); 2-Aminopurine; 7-Dezaa-2,6-Diaminopurine; 7-Dezaa-8-aza-2,6-Diaminopurine; 7-Dezaa-8-aza-2-Aminopurine; 2,6-Diaminopurine; 7-Dezaa-8-aza-adenine; 7-Dezaa-2-Aminopurine; 2-Thiocytosine nucleoside; 3-Methylcytosine nucleoside; 5-Formylcytosine nucleoside; 5-Hydroxymethylcytosine nucleoside; 5-Methylcytosine nucleoside; N4-Acetylcytosine nucleoside; 2′-O-methylcytosine nucleoside; 2′-O-methylcytosine nucleoside; 5,2′-O-dimethylcytosine nucleoside; 5-Formyl-2′-O-methylcytosine nucleoside; Lysidine; N4,2′-O-dimethylcytosine nucleoside; N4-acetyl-2′-O-methylcytosine nucleoside; N4-methylcytosine nucleoside; N4,N4-dimethyl-2′-OMe-cytosine nucleoside TP; 4-methylcytosine nucleoside; 5-aza-cytosine nucleoside; pseudo-iso-cytosine nucleoside; pyrrolo-cytosine nucleoside; α-thio-cytosine nucleoside; 2-(thio)cytosine; 2′-amino-2′-deoxy-CTP; 2′-azido-2′-deoxy-CTP; 2′-deoxy-2′-a-aminocytosine nucleoside TP; 2′-deoxy-2′-a-azido-cytosine nucleoside TP; 3(deaza)5(aza)cytosine; 3(methyl)cytosine; 3-(alkyl)cytosine.3-(deaza)5-(aza)cytosine; 3-(methyl)cytosine nucleoside; 4,2′-O-dimethylcytosine nucleoside; 5-(halo)cytosine; 5-(methyl)cytosine; 5-(propynyl)cytosine; 5-(trifluoromethyl)cytosine; 5-(alkyl)cytosine; 5-(halo)cytosine; 5-(propynyl)cytosine; 5-(trifluoromethyl)cytosine; 5-bromo-cytosine nucleoside; 5-iodo-cytosine nucleoside; 5-propynylcytosine; 6-(azo)cytosine; 6-aza-cytosine nucleoside; aazacytosine; deazacytosine; N4-(acetyl)cytosine; 1-methyl-1-deaza-pseudoisocytosine nucleoside; 1-methyl-pseudoisocytosine nucleoside; 2- Methoxy-5-methyl-cytosine nucleoside; 2-methoxy-cytosine nucleoside; 2-thio-5-methyl-cytosine nucleoside; 4-methoxy-1-methyl-pseudoisocytosine nucleoside; 4-methoxy-pseudoisocytosine nucleoside; 4-thio-1-methyl-1-deaza-pseudoisocytosine nucleoside; 4-thio-1-methyl-pseudoisocytosine nucleoside; 5-aza-zebularine; 5-methyl-zebularine; pyrrolo-pseudoisocytosine nucleoside; zebularine; (E)-5-(2-bromo-vinyl)cytosine nucleoside TP; 2,2′-dehydrated-cytosine nucleoside TP hydrochloride; 2′-fluoro-N4-Bz-cytosine nucleoside TP; 2′-fluoro-N4 -Acetyl-cytosine nucleoside TP; 2′-O-methyl-N4-acetyl-cytosine nucleoside TP; 2′-O-methyl-N4-Bz-cytosine nucleoside TP; 2′-a-ethynylcytosine nucleoside TP; 2′-a-trifluoromethylcytosine nucleoside TP; 2′-b-ethynylcytosine nucleoside TP; 2′-b-trifluoromethylcytosine nucleoside TP; 2′-deoxy-2′,2′-difluorocytosine nucleoside TP; 2′-deoxy-2′-a-mercaptocytosine nucleoside TP; 2′-deoxy-2′-a-thiomethoxycytosine nucleoside TP; 2′-deoxy-2′-b-aminocytosine nucleoside TP; 2′-deoxy-2′-b-azidocytosine nucleoside TP; 2′-deoxy-2′-a-thiomethoxycytosine nucleoside TP; 2′-deoxy-2′-a ... 2′-β-bromocytosine TP; 2′-deoxy-2′-β-chlorocytosine TP; 2′-deoxy-2′-β-fluorocytosine TP; 2′-deoxy-2′-β-iodocytosine TP; 2′-deoxy-2′-β-mercaptocytosine TP; 2′-deoxy-2′-β-thiomethoxycytosine TP; 2′-O-methyl-5-(1-propynyl)cytosine TP; 3′-ethynylcytosine TP; 4′-azidocytosine TP; 4′-carbocycliccytosine TP; 4′-ethynylcytosine TP; 5-(1-propynyl)arsyl-cytosine TP; 5-(2-chloro-phenyl)-2-thiocytosine TP;5-(4-amino-phenyl)-2-thiocytosine nucleoside TP; 5-aminoallyl-CTP; 5-cyanocytosine nucleoside TP; 5-ethynylaza-cytosine nucleoside TP; 5-ethynylcytosine nucleoside TP; 5′-homo-cytosine nucleoside TP; 5-methoxycytosine nucleoside TP; 5-trifluoromethyl-cytosine nucleoside TP; N4-amino-cytosine nucleoside TP; N4-benzylyl-cytosine nucleoside TP; pseudoisocytosine nucleoside; 7-methylguanosine; N2,2′-O-dimethylguanosine; N2-methylguanosine; Wyosine; 1,2′-O-dimethylguanosine; 1-methylguanosine; 2′-O-methylguanosine; 2′-O-ribosylguanosine (phosphate ester); 2′-O-methyl 2′-O-ribosylguanosine (phosphate ester); 7-aminomethyl-7-deazaguanosine; 7-cyano-7-deazaguanosine; Archaeosine; Methylwyoside; N2,7-dimethylguanosine; N2,N2,2′-O-trimethylguanosine; N2,N2,7-trimethylguanosine; N2,N2-dimethylguanosine; N2,7,2′-O-trimethylguanosine; 6-thio-guanosine; 7-deaza-guanosine; 8-sideoxy-guanosine; N1-methyl-guanosine; α-thio-guanosine; 2-(propyl)guanine; 2-(alkyl)guanine; 2′-amino-2′-deoxy-GTP; 2′-azido-2′-deoxy-GTP; 2′-deoxy-2′-a- Aminoguanine TP; 2′-deoxy-2′-a-azidoguanine TP; 6(methyl)guanine; 6-(alkyl)guanine; 6-(methyl)guanine; 6-methyl-guanine; 7(alkyl)guanine; 7(deazido)guanine; 7(methyl)guanine; 7-(alkyl)guanine; 7-(deazido)guanine; 7-(methyl)guanine; 8(alkyl)guanine; 8(alkynyl)guanine; 8(halo)guanine; 8(thioalkyl)guanine; 8-(alkenyl)guanine; 8-(alkyl)guanine; 8-(alkynyl)guanine; 8-(amino)guanine; 8-(halo)guanine; 8-(hydroxy)guanine; 8-(thioalkyl)guanine; 8-(thiol)guanine; azaguanine; deazidoguanine Guanine; N(methyl)guanine; N-(methyl)guanine; 1-methyl-6-thio-guanosine; 6-methoxy-guanosine; 6-thio-7-deaza-8-aza-guanosine; 6-thio-7-deaza-guanosine; 6-thio-7-methyl-guanosine; 7-deaza-8-aza-guanosine; 7-methyl-8-sideoxy-guanosine; N2,N2-di-methyl-6-thio-guanosine; N2-methyl-6-thio-guanosine; 1-Me-GTP; 2′fluoro-N2-isobutyl-guanosine TP; 2′O-methyl-N2-isobutyl-guanosine TP; 2′-a-ethynylguanosine TP; 2′-a-trifluoromethylguanosine TP; 2′-b-ethynylguanosine TP; 2′-b-trifluoromethylguanosine TP;2′-Deoxy-2′,2′-Difluoroguanosine TP; 2′-Deoxy-2′-α-mercaptoguanosine TP; 2′-Deoxy-2′-α-thiomethoxyguanosine TP; 2′-Deoxy-2′-β-aminoguanosine TP; 2′-Deoxy-2′-β-azidoguanosine TP; 2′-Deoxy-2′-β-bromoguanosine TP; 2′-Deoxy-2′-β-chloroguanosine TP; 2′-Deoxy-2′-β-fluoroguanosine TP; 2′-Deoxy-2′-β-iodoguanosine TP; 2′-Deoxy-2′-β-mercaptoguanosine TP; 2′-Deoxy-2′-β-thiomethoxyguanosine TP; 4′-azidoguanosine TP; 4′-carbocyclic guanosine TP; 4′-ethynylguanosine TP; 5′- High-guanosine TP; 8-bromo-guanosine TP; 9-deazaguanosine TP; N2-isobutyl-guanosine TP; 1-methylinosine; inosine; 1,2′-O-dimethylinosine; 2′-O-methylinosine; 7-methylinosine; 2′-O-methylinosine; epoxyqueuosine; galactosylqueuosine; mannosylqueuosine; queuosine; allylamino-thymidine; azathymidine; deazathymidine; deoxy-thymidine; 2′-O-methyluridine; 2-thiouridine; 3-methyluridine; 5-carboxymethyluridine; 5-hydroxyuridine; 5-methyluridine; 5-taurate methyl-2-thiouridine; 5-taurate methyluridine; dihydrouridine; pseudouridine; (3-(3-amino-3- Carboxypropyl)uridine; 1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine; 1-methylpseudouridine; 1-ethylpseudouridine; 2′-O-methyluridine; 2′-O-methylpseudouridine; 2′-O-methyluridine; 2-thio-2′-O-methyluridine; 3-(3-amino-3-carboxypropyl)uridine; 3,2′-O-dimethyluridine; 3-methylpseudo-uridine TP; 4-thiouridine; 5-(carboxyhydroxymethyl)uridine; 5-(carboxyhydroxymethyl)uridine methyl ester; 5,2′-O-dimethyluridine; 5,6-dihydrouridine; 5-aminomethyl-2-thiouridine; 5-carbamoylmethyl-2′-O-methyluridine; 5-carbamoylmethyluridine; 5-carboxyhydroxymethyluridine 5-Carboxyhydroxymethyluridine methyl ester; 5-Carboxymethylaminomethyl-2′-O-methyluridine; 5-Carboxymethylaminomethyl-2-thiouridine; 5-Carboxymethylaminomethyl-2-thiouridine; 5-Carboxymethylaminomethyluridine; 5-Carboxymethylaminomethyluridine; 5-Carboxymethylaminomethyluridine; 5-Carbamoylmethyluridine TP; 5-Methoxycarbonylmethyl-2′-O-methyluridine; 5-Methoxycarbonylmethyl-2-thiouridine; 5-Methoxycarbonylmethyluridine; 5-Methyluridine; 5-Methoxyuridine; 5-Methyl-2-thiouridine; 5-Methoxymethyl-2-selenouridine; 5-Methoxymethyluridine; 5-Methyldihydrouridine; 5-Oxyacetic acid-uridine TP; 5-Oxyacetic acid-methyl ester-uridine TP;N1-Methyl-pseudo-uracil; N1-Ethyl-pseudo-uracil; Uroside 5-oxyacetic acid; Uroside 5-oxyacetic acid methyl ester; 3-(3-amino-3-carboxypropyl)-uridine TP; 5-(isopentenylaminomethyl)-2-thiouridine TP; 5-(isopentenylaminomethyl)-2′-O-methyluridine TP; 5-(isopentenylaminomethyl)uridine TP; 5-propynyluracil; α-Thio-uridine; 1(aminoalkylaminocarbonylvinyl)-2(thio)-pseudo-uracil; 1(aminoalkylaminocarbonylvinyl)-2,4-(dithio)pseudo-uracil; 1(aminoalkylaminocarbonylvinyl)-4(thio)pseudo-uracil; 1(aminoalkylaminocarbonylvinyl)-pseudo-uracil; 1(aminocarbonyl) Vinyl)-2-(thio)-pseudouracil; 1-(aminocarbonylvinyl)-2,4-(dithio)pseudouracil; 1-(aminocarbonylvinyl)-4-(thio)pseudouracil; 1-(aminocarbonylvinyl)-pseudouracil; 1-substituted 2-(thio)-pseudouracil; 1-substituted 2,4-(dithio)pseudouracil; 1-substituted 4-(thio)pseudouracil; 1-substituted pseudouracil; 1-(aminoalkylamino-carbonylvinyl)-2-(thio)-pseudouracil; 1-methyl-3-(3-amino-3-carboxypropyl)pseudouracil TP; 1-methyl-3-(3-amino-3-carboxypropyl)pseudo-UTP; 1-methyl-pseudo-UTP; 1-ethyl-pseudo-UTP; 2-(thio)pseudouracil 2′-Deoxyuridine; 2′-Fluorouracil; 2-(Thio)uracil; 2,4-(Dithio)pseudouracil; 2′-Methyl, 2′-Amino, 2′-Azide, 2′-Fluoro-guanosine; 2′-Amino-2′-Deoxy-UTP; 2′-Azide-2′-Deoxy-UTP; 2′-Azide-Deoxyuridine TP; 2′-O-Methylpseudouridine; 2′-Deoxyuridine; 2′-Fluorouracil; 2′-Deoxy-2′-α-Aminouridine TP; 2′-Deoxy-2′-α-Azide-Uracil TP; 2-Methylpseudouridine; 3(3amino-3-carboxypropyl)uracil; 4(Thio)pseudouracil; 4-(Thio)pseudouracil; 4-(Thio)uracil; 4-Thiouracil; 5(1,3-dithio)uracil; Azolium-1-alkyl)uracil; 5(2-aminopropyl)uracil; 5(aminoalkyl)uracil; 5(dimethylaminoalkyl)uracil; 5(guanidinylalkyl)uracil; 5(methoxycarbonylmethyl)-2-(thio)uracil; 5(methoxycarbonyl-methyl)uracil; 5(methyl)2(thio)uracil; 5(methyl)2,4(dithio)uracil; 5(methyl)4(thio)uracil; 5(methylaminomethyl)-2(thio)uracil; 5(methylaminomethyl)-2,4(dithio)uracil; 5(methylaminomethyl)-4(thio)uracil; 5(propynyl)uracil; 5(trifluoromethyl)uracil; 5-(2-aminopropyl)uracil; 5-(alkyl)-2-(thio)pseudouracil;5-(alkyl)-2,4-(dithio)pseudorazine; 5-(alkyl)-4-(thio)pseudorazine; 5-(alkyl)pseudorazine; 5-(alkyl)uracil; 5-(alkynyl)uracil; 5-(allylamino)uracil; 5-(cyanoalkyl)uracil; 5-(dialkylaminoalkyl)uracil; 5-(dimethylaminoalkyl)uracil; 5-(guanidinylalkyl)uracil; 5-(halo)uracil; 5-(I,3-diazol-I-alkyl)uracil; 5-(methoxy)uracil; 5-(methoxycarbonylmethyl)-2-(thio)uracil; 5-(methoxycarbonyl-methyl)uracil; 5-(methyl)2-(thio)uracil; 5-(methyl)2,4-(dithio)uracil; 5-(methyl) 4-(thio)uracil; 5-(methyl)-2-(thio)pseudorazine; 5-(methyl)-2,4-(dithio)pseudorazine; 5-(methyl)-4-(thio)pseudorazine; 5-(methyl)pseudorazine; 5-(methylaminomethyl)-2-(thio)uracil; 5-(methylaminomethyl)-2,4-(dithio)uracil; 5-(methylaminomethyl)-4-(thio)uracil; 5-(propynyl)uracil; 5-(trifluoromethyl)uracil; 5-aminoallyl-uridine; 5-bromo-uridine; 5-iodo-uridine; 5-uracil; 6-(azo)uracil; 6-(azo)uracil; 6-aza-uridine; allylamino-uracil; azauracil; deazauracil; N3-(methyl) Uracil; pseudo-UTP-1-2-acetic acid; pseudouracil; 4-thio-pseudo-UTP; 1-carboxymethyl-pseudouridine; 1-methyl-1-deaza-pseudouridine; 1-propynyl-uridine; 1-taurate methyl-1-methyl-uridine; 1-taurate methyl-4-thio-uridine; 1-taurate methyl-pseudouridine; 2-methoxy-4-thio-pseudouridine; 2-thio-1-methyl-1-deaza-pseudouridine; 2-thio-1-methyl-pseudouridine; 2-thio-5-aza-uridine; 2-thio-dihydropseudouridine; 2-thio-dihydrouridine; 2-thio-pseudouridine; 4-methoxy-2-thio-pseudouridine; 4-methoxy-pseudouridine; 4-thio-1-methyl-pseudouridine; 4-thio- Pseudouridine; 5-aza-uridine; dihydropseudouridine; (±)1-(2-hydroxypropyl)pseudouridine TP; (2R)-1-(2-hydroxypropyl)pseudouridine TP; (2S)-1-(2-hydroxypropyl)pseudouridine TP; (E)-5-(2-bromo-vinyl)arsyluridine TP; (E)-5-(2-bromo-vinyl)arsyluridine TP; (Z)-5-(2-bromo-vinyl)arsyluridine TP; (Z)-5-(2-bromo-vinyl)uridine TP; 1-(2,2,2-trifluoroethyl)pseudo-UTP; 1-(2,2,3,3,3-pentafluoropropyl)pseudouridine TP; 1-(2,2-diethoxyethyl)pseudouridine TP; 1-(2,4,6-trimethylbenzyl)pseudouridine TP;1-(2,4,6-trimethyl-benzyl) pseudo-UTP; 1-(2,4,6-trimethyl-phenyl) pseudo-UTP; 1-(2-amino-2-carboxyethyl) pseudo-UTP; 1-(2-amino-ethyl) pseudo-UTP; 1-(2-hydroxyethyl) pseudouridine TP; 1-(2-methoxyethyl) pseudouridine TP; 1-(3,4-bis-trifluoromethoxybenzyl) pseudouridine TP; 1-(3,4-dimethoxybenzyl) pseudouridine TP; 1-(3-amino-3-carboxypropyl) pseudo-UTP; 1-(3-amino-propyl) pseudo-UTP; 1-(3-cyclopropyl-prop-2-ynyl) pseudouridine TP; 1-(4-amino-4-carboxybutyl) pseudo-UTP; 1-(4-amino-benzyl) 1-(4-amino-butyl)-UTP; 1-(4-amino-phenyl)-UTP; 1-(4-azidobenzyl)-Uridine TP; 1-(4-bromobenzyl)-Uridine TP; 1-(4-chlorobenzyl)-Uridine TP; 1-(4-fluorobenzyl)-Uridine TP; 1-(4-iodobenzyl)-Uridine TP; 1-(4-methanesulfonylbenzyl)-Uridine TP; 1-(4-methoxybenzyl)-Uridine TP; 1-(4-methoxy-benzyl)-UTP; 1-(4-methoxy-phenyl)-UTP; 1-(4-methylbenzyl)-Uridine TP; 1-(4-methyl-benzyl)-UTP; 1-(4-nitrobenzyl)-Uridine TP; 1-(4-nitro- ... 1-(4-nitro-phenyl)-UTP; 1-(4-thiomethoxybenzyl)-UTP; 1-(4-trifluoromethoxybenzyl)-UTP; 1-(4-trifluoromethylbenzyl)-UTP; 1-(5-amino-pentyl)-UTP; 1-(6-amino-hexyl)-UTP; 1,6-dimethyl-UTP; 1-[3-(2-{2-[2-(2-aminoethoxy)-ethoxy]-ethoxy}-ethoxy)-propionyl]-UTP; 1-{3-[2-(2-aminoethoxy)-ethoxy]-propionyl}-UTP; 1-acetyl-UTP; 1-alkyl-6-(1-propynyl)-UTP; 1-alkyl-6- -(2-Propynyl)-pseudo-UTP; 1-alkyl-6-allyl-pseudo-UTP; 1-alkyl-6-ethynyl-pseudo-UTP; 1-alkyl-6-homallyl-pseudo-UTP; 1-alkyl-6-vinyl-pseudo-UTP; 1-allyl pseudouridine TP; 1-aminomethyl-pseudo-UTP; 1-benzylyl-pseudouridine TP; 1-benzyloxymethyl-pseudouridine TP; 1-benzyl-pseudo-UTP; 1-biotinyl-PEG2-pseudouridine TP; 1-biotinyl-pseudouridine TP; 1-butyl-pseudo-UTP; 1-cyanomethyl-pseudouridine TP; 1-cyclobutylmethyl-pseudo-UTP; 1-cyclobutyl-pseudo-UTP; 1-cycloheptylmethyl-pseudo-UTP; 1-cycloheptyl-pseudo-UTP;1-Cyclohexylmethyl-pseudo-UTP; 1-Cyclohexyl-pseudo-UTP; 1-Cyclooctylmethyl-pseudo-UTP; 1-Cyclooctyl-pseudo-UTP; 1-Cyclopentylmethyl-pseudo-UTP; 1-Cyclopentyl-pseudo-UTP; 1-Cyclopropylmethyl-pseudo-UTP; 1-Cyclopropyl-pseudo-UTP; 1-Ethyl-pseudo-UTP; 1-Hexyl-pseudo-UTP; 1-Homoallyl-pseudouridine TP; 1-Hydroxymethyl-pseudouridine TP; 1-Iso-propyl-pseudo-UTP; 1-Me-2-thio-pseudo-UTP; 1-Me-4-thio-pseudo-UTP; 1-Me-α-thio-pseudo-UTP; 1-Methanesulfonylmethyl-pseudouridine TP; 1-Methoxymethyl-pseudouridine TP; 1-Methyl-6-(2,2,2) -trifluoroethyl) pseudo-UTP; 1-methyl-6-(4-morpholinyl)-pseudo-UTP; 1-methyl-6-(4-thiomorpholinyl)-pseudo-UTP; 1-methyl-6-(substituted phenyl)-pseudo-UTP; 1-methyl-6-amino-pseudo-UTP; 1-methyl-6-azido-pseudo-UTP; 1-methyl-6-bromo-pseudo-UTP; 1-methyl-6-butyl-pseudo-UTP; 1-methyl-6-chloro-pseudo-UTP; 1-methyl-6-cyano-pseudo-UTP; 1-methyl-6-dimethylamino-pseudo-UTP; 1-methyl-6-ethoxy-pseudo-UTP; 1-methyl-6-carboxylic acid ethyl ester-pseudo-UTP; 1-methyl-6-ethyl-pseudo-UTP; 1-methyl-6-fluoro- 1-Methyl-6-formyl-pseudo-UTP; 1-Methyl-6-hydroxyamino-pseudo-UTP; 1-Methyl-6-hydroxy-pseudo-UTP; 1-Methyl-6-iodo-pseudo-UTP; 1-Methyl-6-isopropyl-pseudo-UTP; 1-Methyl-6-methoxy-pseudo-UTP; 1-Methyl-6-methylamino-pseudo-UTP; 1-Methyl-6-phenyl-pseudo-UTP; 1-Methyl-6-propyl-pseudo-UTP; 1-Methyl-6-tert-butyl-pseudo-UTP; 1-Methyl-6-trifluoromethoxy-pseudo-UTP; 1-Methyl-6-trifluoromethyl-pseudo-UTP; 1-morpholinylmethylpseudouridine TP; 1-pentyl-pseudo-UTP; 1-phenyl-pseudo-UTP; 1-trimethyl Acetyl-Pseudouridine TP; 1-Propyno-Pseudouridine TP; 1-Propyno-UTP; 1-Propyno-Pseudouridine; 1-p-Tolyl-Pseudo-UTP; 1-Tertiary-Butyl-Pseudo-UTP; 1-Thiomethoxymethyl-Pseudouridine TP; 1-Thiomorpholinomethyl-Pseudouridine TP; 1-Trifluoroacetyl-Pseudouridine TP; 1-Trifluoromethyl-Pseudo-UTP; 1-Vinyl-Pseudouridine TP; 2,2′-Dehydr-uridine TP; 2′-Bromo-Deoxyuridine TP; 2′-F-5-Methyl-2′-Deoxy-UTP; 2′-OMe-5-MeUTP; 2′-OMe-Pseudo-UTP; 2′-α-Ethynyluridine TP; 2′-α-Trifluoromethyluridine TP; 2′-β-Ethynyluridine TP;2′-β-trifluoromethyluridine TP; 2′-deoxy-2′,2′-difluorouridine TP; 2′-deoxy-2′-α-mercaptouridine TP; 2′-deoxy-2′-α-thiomethoxyuridine TP; 2′-deoxy-2′-β-aminouridine TP; 2′-deoxy-2′-β-azidouridine TP; 2′-deoxy-2′-β-bromouridine TP; 2′-deoxy-2′-β-chlorouridine TP; 2′-deoxy-2′-β-fluorouridine TP; 2′-deoxy-2′-β-iodouridine TP; 2′-deoxy-2′-β-mercaptouridine TP; 2′-deoxy-2′-β-difluorouridine TP; -b-thiomethoxyuridine TP; 2-methoxy-4-thio-uridine; 2-methoxyuridine; 2′-O-methyl-5-(1-propynyl)uridine TP; 3-alkyl-pseudo-UTP; 4′-azidouridine TP; 4′-carbocyclic uridine TP; 4′-ethynyluridine TP; 5-(1-propynyl)arsyluridine TP; 5-(2-furanyl)uridine TP; 5-cyanouridine TP; 5-dimethylaminouridine TP; 5′-homouridine TP; 5-iodo-2′-fluoro-deoxyuridine TP; 5-phenylethynyluridine TP; 5-trideuterylmethyl-6-deuteryluridine TP; 5-trifluoromethyluridine TP; 5-Vinylaradanuridine TP; 6-(2,2,2-trifluoroethyl)-pseudo-UTP; 6-(4-morpholino)-pseudo-UTP; 6-(4-thiomorpholino)-pseudo-UTP; 6-(substituted phenyl)-pseudo-UTP; 6-amino-pseudo-UTP; 6-azido-pseudo-UTP; 6-bromo-pseudo-UTP; 6-butyl-pseudo-UTP; 6-chloro-pseudo-UTP; 6-cyano-pseudo-UTP; 6-dimethylamino-pseudo-UTP; 6-ethoxy-pseudo-UTP; 6-ethylcarboxylate-pseudo-UTP; 6-ethyl-pseudo-UTP; 6-fluoro-pseudo-UTP; 6-methyl Acyl-pseudo-UTP; 6-hydroxyamino-pseudo-UTP; 6-hydroxy-pseudo-UTP; 6-iodo-pseudo-UTP; 6-iso-propyl-pseudo-UTP; 6-methoxy-pseudo-UTP; 6-methylamino-pseudo-UTP; 6-methyl-pseudo-UTP; 6-phenyl-pseudo-UTP; 6-phenyl-pseudo-UTP; 6-propyl-pseudo-UTP; 6-tert-butyl-pseudo-UTP; 6-trifluoromethoxy-pseudo-UTP; 6-trifluoromethyl-pseudo-UTP; α-thio-pseudo-UTP; pseudouridine 1-(4-methylbenzenesulfonic acid)TP; pseudouridine 1-(4-methylbenzoic acid)TP; pseudouridine TP 1-[3-(2-ethoxy)]propionic acid; pseudouridine TP 1-[3-{2-(2-[2-(2-(2-ethoxy)-ethoxy]-ethoxy)-ethoxy}]propionic acid; pseudouridine TP 1-[3-{2-(2-[2-{2(2-ethoxy)-ethoxy}-ethoxy]-ethoxy)-ethoxy}]propionic acid;Pseudoruridine TP 1-[3-{2-(2-[2-ethoxy]-ethoxy)-ethoxy}]propionic acid; Pseudoruridine TP 1-[3-{2-(2-ethoxy)-ethoxy}]propionic acid; Pseudoruridine TP 1-methylphosphonic acid; Pseudoruridine TP Diethyl 1-methylphosphonate; pseudo-UTP-N1-3-propionic acid; pseudo-UTP-N1-4-butyric acid; pseudo-UTP-N1-5-valeric acid; pseudo-UTP-N1-6-hexanoic acid; pseudo-UTP-N1-7-heptanoic acid; pseudo-UTP-N1-methyl-terebenzoic acid; pseudo-UTP-N1-terebenzoic acid; Wybutosine; hydroxywybutosine; isowybutosine; peroxywybutosine; incompletely modified hydroxywybutosine; 4-demethylwybutosine; 2,6-(diamino)purine; 1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl; 1,3-(diaza)-2-(side-oxy) )-Phenylothiazine-I-yl; 1,3-(diaza)-2-(sideoxy)-phenoxazine-1-yl; 1,3,5-(triaza)-2,6-(dioxa)-naphthalene; 2(amino)purine; 2,4,5-(trimethyl)phenyl; 2′methyl, 2′amino, 2′azido, 2′fluoro-cytosine nucleoside; 2′methyl, 2′amino, 2′azido, 2′fluoro-adenine; 2′methyl, 2′amino, 2′azido, 2′fluoro-uridine; 2′-amino-2′-deoxyribose; 2-amino-6-chloro-purine; 2-aza-inosine; 2′-azido-2′-deoxyribose; 2′fluoro-2′-deoxyribose; via 2′ -Fluoro-modified bases; 2′-O-methyl-ribose; 2-side-oxy-7-aminopyridinidin-3-yl; 2-side-oxy-pyridinidin-3-yl; 2-pyridinone; 3-nitropyrrole; 3-(methyl)-7-(propynyl)isoquinolone; 3-(methyl)isoquinolone; 4-(fluoro)-6-(methyl)benzimidazole; 4-(methyl)benzimidazole; 4-(methyl)indolyl; 4,6-(dimethyl)indolyl; 5-nitroindol; 5-substituted pyrimidine; 5-(methyl)isoquinolone; 5-nitroindol; 6-(aza)pyrimidine; 6-(azo)thymine; 6-(methyl)-7-(aza)indolyl; 6-chloro- Purine; 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; 7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl; 7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl; 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(sideoxy)-phenoxazine-1-yl; 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(sideoxy)-phenoxazine-1-yl; 7-(aza)indolyl;7-(guanidinoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-I-yl; 7-(guanidinoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-I-yl; 7-(guanidinoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazine-1-yl; 7-(guanidinoalkylhydroxy)-1,3-(diaza)-2-(sideoxy)-phenoxazine-1-yl; 7-(guanidinoalkylhydroxy)-I,3-(diaza)-2-(sideoxy)-phenoxazine-I-yl; 7-(guanidinoalkylhydroxy)-I,3-(diaza)-2-(sideoxy)-phenoxazine-I-yl ; 7-(propynyl)isoquinolone; 7-(propynyl)isoquinolone; propynyl-7-(aza)indolyl; 7-deaza-inosine; 7-substituted 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 7-substituted 1,3-(diaza)-2-(sideoxy)-phenoxazin-1-yl; 9-(methyl)-imidazopyridyl; aminoindolyl; anthracene; bis-o-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; bis-o-substituted 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; difluorotolyl; hypoxanthine; imidazopyridyl; inosine; isoquinolone; isoguanosine; N 2-Substituted purines; N6-methyl-2-amino-purines; N6-substituted purines; N-alkylated derivatives; naphthyl; nitrobenzimidazolyl; nitroimidazolyl; nitroindazolyl; nitropyrazolyl; nubularine; O6-substituted purines; O-alkylated derivatives; o-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; o-substituted 6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; Oxoformycin TP; p-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl; para-substituted 6-phenyl-pyrrolo-pyrimidin -2-keto-3-yl; pentaphenyl; benzanthyl; phenyl; propynyl-7-(aza)indolyl; pyrene; pyridopyrimidine-3-yl; pyridopyrimidine-3-yl; 2-sideoxy-7-amino-pyridopyrimidine-3-yl; pyrrolo-pyrimidine-2-keto-3-yl; pyrrolo-pyrimidineyl; pyrrolo-pyrazinyl; stilbene; substituted 1,2,4-triazole; tetraphenyl; tubercidine; xanthine; xanthine nucleoside-5′-TP; 2-thio-zebraline; 5-aza-2-thio-zebraline; 7-deaza-2-amino-purine; pyridin-4-ketoribonucleoside; 2-amino-riboside-TP; mesomycin A TP; mesomycin B TP; pyrrolosine TP; 2′-OH-arabino-adenosine TP; 2′-OH-arabino-cytosine TP; 2′-OH-arabino-uridine TP;2′-OH-arabinoguanosine TP; 5-(2-methoxycarbonylvinyl)uridine TP; and N6-(19-amino-pentaenolide)adenosine TP.

[0222] In one embodiment, the vaccine is a protein vaccine. The protein vaccine includes the gE protein antigen variant described above.

[0223] Furthermore, the protein vaccine is carried by a carrier comprising serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid, diphtheria toxoid, genetically modified cross-reactive substances of diphtheria toxoid, CRM197, meningococcal outer membrane protein complex and Haemophilus influenzae protein D, rEPA, keyhole hemocyanin, and / or flagellin.

[0224] Furthermore, the protein vaccine is presented by a variety of antigen presentation systems.

[0225] Furthermore, the antigen presentation system includes one or more of the following: a Lys-based dendritic architecture, helper T cell epitopes, immunostimulatory lipophilic moieties, cell-penetrating peptides, free radical-induced polymerization, self-assembled nanoparticles, and gold nanoparticles.

[0226] In one embodiment, the vaccine is a viral vector vaccine. The viral vector vaccine comprises a nucleotide sequence encoding an open reading frame of at least one of the aforementioned gE protein antigen variants.

[0227] Furthermore, the nucleotide sequence is codon-optimized or codon-degenerate.

[0228] Furthermore, the nucleotide sequence is codon-selected as a change from the natural type. Any suitable viral vector can be used for the construction of the vaccine of the present invention, and suitable viral vectors have been described in Dicks et al. (Vaccine. 25 Feb 2015; 33(9):1121-8. doi:10.1016 / j.vaccine.2015.01.042.Epub 25 Jan 2015), Antrobus et al. (Mol Ther. 2014 Mar; 22(3):668-74. doi:10.1038 / mt.2013.284.Epub 30 Dec 2013), and Warimwe et al. (Virol J. 5 Dec 2013; 10:349. doi:10.1186 / 1743-422X-10-349), which are incorporated herein by reference.

[0229] Furthermore, the vector in the viral vector vaccine self-assembles into virus-like particles.

[0230] Furthermore, the carrier includes a self-deactivating carrier.

[0231] Furthermore, the virus is selected from lentiviruses, influenza viruses, hepatitis viruses, alpha viruses, filoviruses, adenoviruses, adeno-associated viruses, and / or flaviviruses.

[0232] Furthermore, the virus is an adenovirus or adeno-associated virus.

[0233] Furthermore, the virus is an adenovirus; for example, a human, simian, or chimpanzee adenovirus. In one embodiment, the viral vector comprises an adenovirus, such as group E simian adenovirus. The viral vector may contain ChAdOx1 or ChAdOx2. The viral vector may contain AdCh63. The viral vector may contain AdC3 or AdH6. In one embodiment, the viral vector is a human serotype.

[0234] In some embodiments of the present invention, the vaccine further includes gI, gB, gH, gK, gL, gC, gN, and / or gM glycoproteins of varicella-zoster virus. In some embodiments, the vaccine may be any combination of VZV gE with at least one of the following peptides: gI, gB, gH, gK, gL, gC, gN, and gM, or immunogenic fragments or epitopes thereof. In some embodiments, the vaccine may be any combination of VZV gE, gI, and at least one of the following peptides: gB, gH, gK, gL, gC, gN, and gM, or immunogenic fragments or epitopes thereof.

[0235] In embodiments of the present invention, the vaccine further includes pharmaceutically acceptable adjuvants.

[0236] Furthermore, the pharmaceutically acceptable excipients include those required for intravenous infusion, subcutaneous injection, intravenous bolus injection, intravitreal injection, and intramuscular injection.

[0237] Furthermore, the adjuvants include physiological saline, solution tension regulators, solution osmotic pressure regulators, viscosity-reducing adjuvants, buffer solutions, or surfactants.

[0238] Furthermore, the solution tension regulator includes sodium chloride or arginine.

[0239] Furthermore, the solution osmotic pressure regulator includes sucrose, trehalose, sorbitol, or mannitol.

[0240] Furthermore, the viscosity-reducing excipients include arginine, sodium chloride, glycine, proline, or lysine.

[0241] Furthermore, the buffer solution includes histidine, phosphoric acid, citric acid, acetate, succinic acid, lactic acid, tromethamine, aspartic acid, glutamic acid, adipic acid, or MES.

[0242] Furthermore, the surfactant includes polysorbate 20, polysorbate 80, poloxamer 188, or polyethylene glycol 3350.

[0243] The term "pharmaceutically acceptable" refers to a carrier that is generally chemically and / or physically compatible with the other components constituting the formulation and physiologically compatible with its recipient. Pharmaceutically acceptable excipients used in the vaccine compositions of the present invention may include, but are not limited to, pharmaceutically acceptable liquid, gel, or solid carriers, aqueous mediators (e.g., sodium chloride injection, Ringer's solution, isotonic glucose injection, sterile water injection, or Ringer's glucose and lactate injection), non-aqueous mediators (e.g., non-volatile plant oils, cottonseed oil, corn oil, sesame oil, or peanut oil), antimicrobial agents, isotonic agents (e.g., sodium chloride or dextrose), buffers (e.g., phosphate or citrate buffers), antioxidants (e.g., sodium bisulfate), anesthetics (e.g., procaine hydrochloride), suspending / dispersing agents (e.g., sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, or polyvinylpyrrolidone), chelating agents (e.g., EDTA (ethylenediaminetetraacetic acid) or EGTA (ethylene glycol tetraacetic acid)), emulsifiers (e.g., polysorbate 80 (Tween 80)), diluents, adjuvants, excipients, or non-toxic excipients, other components known in the art, or various combinations thereof. Suitable components may include, for example, fillers, binders, disintegrants, buffers, preservatives, lubricants, flavorings, thickeners, colorants, or emulsifiers.

[0244] In embodiments of the present invention, the term "vaccine" is defined as a composition for inducing or stimulating an immune response against an antigen or fragment thereof within the composition to protect or treat an organism from disease.

[0245] The present invention provides a derivative of a gE protein antigen variant, the derivative comprising a complex formed by direct or indirect coupling of the aforementioned gE protein antigen variant to a detectable marker.

[0246] Furthermore, the detectable markers include fluorescent dyes, chemiluminescent compounds, radioactive isotopes, electron-dense reagents, enzymes, colored particles, or biotin.

[0247] In this invention, the term "derivative" can be a nucleic acid molecule, as a DNA molecule, encoding a polynucleotide as defined above, or a nucleic acid molecule containing a polynucleotide as defined above, or a polynucleotide with a complementary sequence. In the context of this invention, the term "derivative" also refers to a longer or shorter polynucleotide and / or polypeptide having, for example, at least 40%, 50%, 60%, 65%, 70%, or 75%, more preferably at least 85%, for example at least 90%, and even more preferably at least 95% or 100% identity percentage with the mentioned sequence or its complementary sequence or its DNA or RNA corresponding sequence. The term "derivative" also includes modified synthetic oligonucleotides. The term "derivative" may also include nucleotide analogs, i.e., naturally occurring ribonucleotides or deoxyribonucleotides substituted with non-naturally occurring nucleotides. The term "derivative" also includes nucleic acids or polypeptides that can be produced by mutating one or more nucleotides or amino acids in their sequence, equivalents, or precursor sequences. The term "derivative" also includes at least one functional fragment of a polynucleotide.

[0248] The term "detectable marker" refers to a reagent that is detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Useful detectable markers include, but are not limited to, fluorescent dyes, chemiluminescent compounds, radioactive isotopes, electron-dense reagents, enzymes, colored particles, and biotin. Detectable markers often produce measurable signals, such as radioactivity, fluorescence, color, or enzyme activity. Antibodies conjugated to detectable reagents can be used for diagnostic or therapeutic purposes. Examples of detectable reagents include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals using various positron emission tomography techniques, and non-radioactive paramagnetic metal ions. Detectable substances can be directly linked to or conjugated to antibodies, or indirectly through intermediates such as linkers known in the art, using techniques known in the art. See U.S. Patent No. 4,741,900, which describes the conjugation of metal ions to antibodies for diagnostic purposes. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, and acetylcholinesterase; examples of suitable cofactor complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, luciferin, luciferin isothiocyanate, rhodamine, dichlorotriazine luciferin, dansyl chloride, and phycoerythrin; one example of a luminescent material is luminescent ammonia; examples of bioluminescent materials include insect luciferin and luminescent proteins.

[0249] This invention provides the application of the aforementioned gE protein antigen variant in the preparation of products for detecting varicella-zoster virus.

[0250] This invention provides the use of the aforementioned gE protein antigen variants in the preparation, capture, or screening of antibodies that recognize the varicella-zoster virus gE protein.

[0251] The term "antibody" refers to a heterotetraglycoprotein of approximately 150,000 Daltons with similar structural features, composed of two identical light chains (L) and two identical heavy chains (H). Each light chain is linked to the heavy chain by a covalent disulfide bond, although the number of disulfide bonds between heavy chains varies among different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bonds. Each heavy chain has a variable region (VH) at one end, followed by multiple constant regions. Each light chain has a variable region (VL) at one end and a constant region at the other; the constant region of the light chain is opposite the first constant region of the heavy chain, and the variable region of the light chain is opposite the variable region of the heavy chain. Specific amino acid residues form interfaces between the variable regions of the light and heavy chains.

[0252] This invention provides the use of the aforementioned gE protein antigen variants, the aforementioned nucleic acid molecules, the aforementioned vectors, the aforementioned cells, the aforementioned pharmaceutical compositions, or the aforementioned vaccines in the preparation of products for the treatment or prevention of varicella-zoster virus-related diseases.

[0253] This invention provides the use of the aforementioned gE protein antigen variant, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned cell, the aforementioned pharmaceutical composition, or the aforementioned vaccine in the preparation of products that inhibit varicella-zoster virus.

[0254] This invention provides the use of the aforementioned gE protein antigen variants, the aforementioned nucleic acid molecules, the aforementioned vectors, the aforementioned cells, the aforementioned pharmaceutical compositions, or the aforementioned vaccines in the preparation of antibodies against the gE protein.

[0255] This invention provides a method for screening, detecting, or isolating varicella-zoster virus gE protein antibodies, the method comprising screening, detecting, or isolating varicella-zoster virus gE protein antibodies using the aforementioned gE protein antigen variants.

[0256] This invention provides a method for producing the aforementioned gE protein antigen variant, the method comprising the following steps:

[0257] (i) Transformed into host cells using the vector described above, or directly using the cells described above;

[0258] (ii) Culturing the host cells or cells from step (i) under suitable conditions;

[0259] (iii) The gE protein antigen variant described above was isolated and purified from the host cell culture medium.

[0260] The present invention provides a detection reagent, a test strip, and a kit comprising the thereof, which contains the aforementioned gE protein antigen variant, the aforementioned nucleic acid molecule, the aforementioned carrier, the aforementioned cell, the aforementioned pharmaceutical composition, or the aforementioned derivative.

[0261] A method for inhibiting or reducing varicella-zoster virus in subjects who have varicella-zoster virus-associated disease or are at risk of developing varicella-zoster virus-associated disease, comprising administering the aforementioned vaccine to the subject.

[0262] A method for inducing an immune response in a subject, the method comprising delivering, in an amount sufficient to induce an immune response in humans, the aforementioned gE protein antigen variant, the aforementioned nucleic acid molecule, the aforementioned vector, the aforementioned pharmaceutical composition, and / or the aforementioned vaccine.

[0263] Furthermore, the immune response includes an antigen-specific immune response.

[0264] Furthermore, the antigen-specific immune response includes a T-cell response or a B-cell response.

[0265] Furthermore, the subject was given a single dose of the aforementioned vaccine.

[0266] Furthermore, the subject was administered the first and second doses (i.e., booster doses) of the previously described vaccine.

[0267] Furthermore, the method administers the vaccine to the subject via intravenous drip, subcutaneous injection, intravenous bolus injection, intravitreal injection, or intramuscular injection.

[0268] A method for preventing or treating varicella-zoster virus infection, the method comprising administering the aforementioned vaccine to a subject.

[0269] The beneficial effects of this invention are:

[0270] This invention relates to an antigenic variant of the varicella-zoster virus gE protein, which exhibits high expression levels and high immunogenicity. Therefore, when this antigenic variant is used as a vaccine composition, it demonstrates superior safety compared to a live attenuated vaccine; and compared to the antigen of a recombinant protein vaccine, it exhibits better immunogenicity due to its high expression level on the cell membrane surface. It can be designed as a safer viral vector vaccine or a more immunogenic RNA vaccine for the prevention of shingles. Attached Figure Description

[0271] Figure 1 This is a flow cytometry graph showing the expression of gE on the cell membrane surface;

[0272] Figure 2 This is a Western blot graph showing gE expression.

[0273] Figure 3 This is a graph showing the results of mRNA integrity detection using agarose gel electrophoresis.

[0274] Figure 4 This is a flow cytometry graph showing the expression levels of different antigen variants on the cell membrane surface.

[0275] Figure 5 This is a graph showing the results of the immune assessment.

[0276] Figure 6 This is a graph showing the results of testing the immunization efficacy of the mRNA-gE truncated vaccine compared to other vaccines;

[0277] Figure 7 This is a graph showing the results of testing the immunization efficacy of the adenovirus gE truncated vaccine compared to other vaccines. Detailed Implementation

[0278] The present invention will be further described below with reference to embodiments. The following description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make equivalent modifications to the disclosed technical content to create equivalent embodiments. Any simple modifications or equivalent changes made to the following embodiments based on the technical essence of the present invention without departing from the scope of the invention are all within the protection scope of the present invention.

[0279] Example 1

[0280] 1. Construction of gE antigen variants

[0281] The C-terminus of the gE antigen was systematically truncated, and the desired gE fragments were obtained by PCR. Each gE fragment was then digested with a restriction endonuclease and inserted into the pCAGGS vector. After sequencing confirmation, the DNA of the pCAGGS vector containing different variant gE fragments was obtained using a plasmid extraction kit.

[0282] The amino acid sequences of the gE antigen variants are shown in Table 1.

[0283] Table 1

[0284] 546 The carboxyl terminus of the 546th amino acid residue is truncated. GSK vaccine sequence 547 The carboxyl terminus of the 547th amino acid residue is truncated. SEQ ID NO:2 551 The carboxyl terminus of the 551st amino acid residue is truncated. SEQ ID NO:3 556 The carboxyl terminus of the 556th amino acid residue is truncated. SEQ ID NO:4 561 The carboxyl terminus of the 561st amino acid residue is truncated. SEQ ID NO:5 566 The carboxyl terminus of the 566th amino acid residue is truncated. SEQ ID NO:6 571 The carboxyl terminus of the 571st amino acid residue is truncated. SEQ ID NO:7 576 The carboxyl terminus of the 576th amino acid residue is truncated. SEQ ID NO:8 581 The carboxyl terminus of the 581st amino acid residue is truncated. SEQ ID NO:9 586 The carboxyl terminus of the 586th amino acid residue is truncated. SEQ ID NO:10 591 The carboxyl terminus of the 591st amino acid residue is truncated. SEQ ID NO:11 596 The carboxyl terminus of the 596th amino acid residue is truncated. SEQ ID NO:12 601 The carboxyl terminus of the 601st amino acid residue is truncated. SEQ ID NO:13 606 The carboxyl terminus of the 606th amino acid residue is truncated. SEQ ID NO:14 611 The carboxyl terminus of the 611th amino acid residue is truncated. SEQ ID NO:15 616 The carboxyl terminus of the 616th amino acid residue is truncated. SEQ ID NO:16 621 The carboxyl terminus of the 621st amino acid residue is truncated. SEQ ID NO:17 623 The carboxyl terminus of amino acid residue 623 is truncated. SEQ ID NO:18

[0285] 2. Transient transfection of 293T cells

[0286] Seed cells one day before transfection, aiming for a cell density of 70%–80% at transfection. For each well, dilute 2 μg of target DNA with 100 μL of serum-free medium and mix thoroughly to prepare a DNA dilution buffer. Immediately add 4 μL of PEI 40000 transfection reagent to 100 μL of DNA dilution buffer, vortex for 10 seconds, and mix thoroughly. Incubate at room temperature for 10–15 min to form the DNA-PEI cationic nucleic acid transfection reagent complex. Directly add 100 μL of the DNA-PEI nucleic acid-PEI complex to the cells, shake the culture plate, and gently mix. Incubate at 37°C in a 5% CO2 incubator for 36 h.

[0287] 3. Flow cytometry detection of gE expression on cell membrane surface

[0288] Take 1×10 6 Add 50 μl of primary antibody (gE extracellular protein mouse immune serum) to the cell suspension, resuspend, and incubate on ice in the dark for 30 minutes. After centrifugation, resuspend the cells twice with 200 μl of cell buffer. Resuspend the pellet in 50 μl of cell buffer containing secondary antibody (FITC anti-mouse IgG Antibody) and incubate on ice in the dark for 30 minutes. After centrifugation, resuspend the cells twice with 200 μl of cell buffer. Resuspend in 200 μl of cell buffer and perform flow cytometry analysis.

[0289] Flow cytometry analysis results as follows Figure 1 As shown, the results indicate that there is no binding between 546 / 547 truncations, and the proportion of 551 truncations is significantly higher than the others. Overall, 591 truncation is the dividing point, and the proportion of truncations before 591 is generally higher than that after 591.

[0290] 4. Western blot detection of gE expression

[0291] After harvesting the cells, centrifuge at 350xg for 5 minutes, resuspend in PBS, centrifuge again, and then resuspend the intracellular sample in 200 μl of RIPA lysis buffer. Perform Western blotting on the intracellular sample to determine its correct expression.

[0292] The test was performed using laboratory-prepared gE protein mouse immune serum as the primary antibody, and the results are as follows: Figure 2 As shown in the figure, the results indicate that the target band was detected in all truncated forms.

[0293] 5. mRNA preparation

[0294] mRNA transcription template preparation

[0295] Different antigenic variant sequences of VZV gE nucleotides were added with T7 promoter, 5'UTR, and 3'UTR sequences. The resulting DNA sequences were cloned into the pUC57 vector to obtain mRNA transcription templates. After successful sequencing, plasmids were extracted. The transcription template plasmids were then linearized by restriction endonuclease Hind III, and the linearized transcription templates were recovered using DNA magnetic beads.

[0296] mRNA stock solution preparation

[0297] mRNA transcription was performed using the Vazyme T7 High Yield RNA Transcription Kit (N1-Me-Pseudo UTP) under the following conditions: incubation at 37°C for 2 hours. After transcription, 1 μl of DNase I was added to the reaction system and incubated at 37°C for 15 minutes to digest the transcribed DNA template. The transcribed RNA was purified using VAHTS RNA purification magnetic beads, and the yield of in vitro transcribed RNA was determined by measuring the A260 reading of the product. The higher-order structure of the 5' end of the RNA was opened by heat denaturation (heating at 65°C for 5 minutes, then placing on ice for 5 minutes), and then the transcribed mRNA was capped using Vaccinia Capping Enzyme and 2'-O-Methyltransferase to form a cap1 cap. A polyadenylated tail was then added to the 3' end of the mRNA using E. coli Poly(A) Polymerase+. Magnetic bead purification was performed after both capping and tailing; the purified sample after tailing is the mRNA stock solution.

[0298] 6. Detection of mRNA stock solution

[0299] The integrity of mRNA was detected using Urea-TBE agarose gel electrophoresis, and the results are as follows: Figure 3 As shown in the figure, the results indicate that the mRNA was of good integrity and the bands were single.

[0300] Flow cytometry was used to determine the expression levels of different antigen variants on the cell membrane surface. 293T cells were transfected with different truncated gE mRNAs, and surface flow cytometry was performed 36 hours later. The primary antibody was V5 protein 3 immune serum, and the secondary antibody was FITC anti-mouse IgG antibody.

[0301] The results are as follows Figure 4 As shown in the figure, the results indicate that gE-mRNAs of different truncated lengths were successfully expressed after transfection, and the trend of mRNA transfection results was basically consistent with that of plasmid transfection results.

[0302] 7. mRNA-LNP preparation

[0303] mRNA was packaged into nanoparticles using microfluidic technology. The aqueous phase contained mRNA at a concentration of 150 μg / ml (50 mM sodium acetate buffer, pH 4.0). The ethanol phase was a lipid mixture prepared from ALC-0315, ALC-0159, DSPC, and cholesterol in a molar ratio of 46.3:9.4:42.7:1.6. The packaging was performed using a Myanna INanoL instrument. The total flow rate of the aqueous and ethanol phases was 12 ml / min, with a volume ratio of 1:3 and a flow rate ratio of 1:3. After packaging, the mRNA-gE nanoparticles were replaced with PBS using a 30 kDa ultrafiltration concentrator to obtain mRNA-gE-LNP, the vaccine composition.

[0304] 8. Testing of vaccine compositions

[0305] The particle size, polydispersity index (PDI), and zeta potential of each obtained mRNA vaccine composition were measured using a Zetasizer Nano ZS nanoparticle analyzer. The LNP nanoparticle vaccines were incubated in 2% Triton-TE buffer at room temperature for 10 min. Nucleic acid concentration was measured and nucleic acid loading efficiency was calculated using the Quant-iT RiboGreen RNA Reagent Kit.

[0306] The physicochemical properties of VZV with different truncated gE mRNA-LNPs, such as particle size, particle size distribution, zeta potential, and encapsulation efficiency, were detected. The particle size was around 100 nm; the PDI was less than 0.2; the zeta potential was around 0 mV; and the encapsulation efficiency was 90%, which met the requirements for immunoassay.

[0307] 9. Animal immunization

[0308] The purified mRNA-gE-LNP was adjusted to a concentration of 100 μg / ml with PBS and used to immunize Balb / c mice. The immunization program was as follows: mice in each group were intramuscularly injected at weeks 0 and 3, with an immunization dose of 100 μl, approximately 5 μg per mouse. Blood samples were collected from the orbital sinus at weeks 2 and 5 post-immunization to assess humoral immunity levels; at week 5, mice were sacrificed and spleen lymphocytes were collected to assess cellular immunity levels.

[0309] 10. Immune assessment

[0310] Humoral immunity assessment

[0311] The gE extracellular domain protein was used as the coating protein, with a coating amount of 50 ng / well, and coating was performed overnight at 4°C. Serum samples were serially diluted at 2 and 5 weeks and then analyzed by ELISA to assess binding activity. The fitted curves were summarized and analyzed using GraphPad Prism.

[0312] Cellular immune assessment

[0313] After isolating splenic lymphocytes, the concentration of splenic lymphocytes was adjusted to 1.2 × 10⁻⁶. 6 Cells / 200 μl were incubated overnight in a CO2 incubator at 37°C. After centrifugation, cells were resuspended in 50 μl of culture medium, and 50 μl of 4 μg / mL peptide library working solution, 5 μg / mL co-stimulating antibody CD28, and 5 μg / mL co-stimulating antibody CD49d were added. The mixture was incubated for 1 hour, and then 5 μg / mL brefeldin A was added. The cells were incubated under the same conditions for 5 hours to prevent the release of cytokines. Cells were washed twice with 200 μl of buffer, and 2.5 μg / mL CD16 / 32 antibody was added. The cells were incubated at 4°C in the dark for 10 minutes to inhibit the non-specific binding of Fc. Cells were washed twice with 200 μl of buffer, and 100 μl of Zombie NIR dye (1:500) was used to resuspend the cells. The cells were incubated at room temperature in the dark for 20 minutes. Wash cells twice with 200 μl buffer, add 1.25 μg / mL FITC anti-mouse CD4 Antibody and 5 μg / mL PerCP / Cyanine 5.5 anti-mouse CD8a Antibody, in a 100 μl solution, and incubate at 4°C in the dark for 30 min. After fixation and membrane perforation, add 1.25 μg / mL APC anti-mouse IL-2 Antibody, 1.25 μg / mL PE anti-mouse IFN-γ antibody, and 1.25 μg / mL Brilliant Violet 421. TM Anti-mouse TNF-α antibody, 100 μl solution, incubated at room temperature in the dark for 30 min. After washing twice, the cells were resuspended in 200 μl, and viable cells were harvested for 10 weeks. Flow cytometry analysis was performed on the resulting 10 weeks of activity.

[0314] The results are as follows Figure 5 As shown, the results indicated that: (1) Strong antibody levels were elicited in both groups after 2 immunizations (5 weeks), with the antibody levels of mRNA-571 and 591 being significantly higher than those of mRNA-623. (2) All mRNA groups significantly induced antigen-specific functional T cell immune responses, including CD4+ and CD8+ T cell immune responses, and these responses were significantly higher than those in the control group. (3) The level of CD4 T cells secreting IFN-γ induced in mRNA-571 mice was significantly higher than that in mRNA-623 mice, and the level of CD8 T cells secreting IFN-γ induced in mRNA-591 mice was significantly higher than that in mRNA-623 mice.

[0315] 11. Comparison of mRNA-gE truncated vaccines with other vaccines

[0316] Balb / c and C57BL / 6 mice were immunized with mRNA-571, mRNA-591, shingrix, and HZV-live, respectively. The immunization schedule was as follows: 5 mice per group received intramuscular injections at weeks 0 and 3. The mRNA vaccine dose per mouse was 10 μg; the shingrix vaccine and the live attenuated HZV-live vaccine dose per mouse were one-tenth of the vaccine dose, with the live attenuated HZV-live vaccine administered only once at week 3. Five weeks post-immunization, mice were sacrificed, and blood was collected to assess humoral immunity levels. Simultaneously, spleen lymphocytes were collected to assess cellular immunity levels.

[0317] The results are as follows Figure 6 As shown, the results indicate that: (1) the antibody response trend was consistent in both strains, and the induced responses of mRNA vaccines 571 and 591 were comparable, superior to shingrix and the live attenuated vaccine HZV-live. (2) in mice of both strains, mRNA vaccines 571 and 591 significantly induced antigen-specific functional T-cell immune responses (including CD4+ and CD8+ T-cell immune responses), and were significantly superior to shingrix and the live attenuated vaccine HZV-live. (3) the CD4 T-cell levels of IL-2 induced in mice immunized with mRNA vaccines 571 and 591 were significantly higher than those immunized with shingrix and the live attenuated vaccine HZV-live, suggesting that they can induce long-term immunity.

[0318] 12. Comparison of adenovirus gE truncated vaccine and other vaccines

[0319] Adenovirus vaccines were prepared using a truncated gE-573 sequence. C57BL / 6 mice were immunized with adenovirus Ad-573, shingrix, and HZV-live.

[0320] The immunization schedule was as follows: Five mice per group received intramuscular injections at weeks 0 and 3. The adenovirus vaccine dose per mouse was 10^10 VP; the shingrix vaccine and the live attenuated vaccine HZV-live were administered at one-tenth of the vaccine dose per mouse. Five weeks post-immunization, mice were sacrificed, and blood samples were collected to assess humoral immunity levels. Simultaneously, spleen lymphocytes were harvested to assess cellular immunity levels.

[0321] The results are as follows Figure 7 As shown, the results indicate that the levels of CD4T cells secreting IFN-γ induced in mice immunized with Ad-573 and Shingrix were comparable.

[0322] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. A variant of the gE protein antigen, characterized in that, The gE protein antigen variant is a polypeptide truncated at the carboxyl terminus of amino acid residue 571, the position of which is referenced in the amino acid sequence SEQ ID NO:1; The amino acid sequence of the gE protein antigen variant is shown in SEQ ID NO:

7.

2. A nucleic acid molecule, characterized in that, The nucleic acid molecule encodes the gE protein antigen variant of claim 1.

3. A carrier, characterized in that, The carrier comprises the nucleic acid molecule as described in claim 2.

4. The carrier according to claim 3, characterized in that, The vector is selected from plasmid vectors, lentiviral vectors, adenovirus vectors, adeno-associated virus vectors, piggyBac vectors, or Sleeping Beauty transposable vectors.

5. A cell, characterized in that, The cell comprises the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, and / or the vector of claim 3 or 4.

6. The cell according to claim 5, characterized in that, The cells are selected from prokaryotic or eukaryotic cells.

7. The cell according to claim 6, characterized in that, The eukaryotic cells were 293T cells.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, and / or the cell of any one of claims 5-7.

9. A vaccine, characterized in that, The active ingredient in the vaccine is an RNA polynucleotide encoding an open reading frame of the gE protein antigen variant of claim 1, and / or the gE protein antigen variant of claim 1.

10. The vaccine according to claim 9, characterized in that, The vaccine is selected from RNA vaccines, protein vaccines, or viral vector vaccines.

11. The vaccine according to claim 10, characterized in that, The RNA vaccine comprises an RNA polynucleotide encoding an open reading frame of the gE protein antigen variant of claim 1.

12. The vaccine according to claim 11, characterized in that, The open reading frames of the RNA polynucleotides are codon-optimized.

13. The vaccine according to claim 11, characterized in that, The RNA polynucleotide contains at least one chemical modification.

14. The vaccine according to claim 13, characterized in that, The chemical modification is selected from any group of the following: pseudouridine, N1-methylpseuuridine, N1-ethylpseuuridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseuuridine, 2-thio-1-methylpseuuridine, 2-thio-5-aza-uridine, 2-thio-dihydropseuuridine, 2-thio-dihydrouridine, 2-thio-pseuuridine, 4-methoxy-2-thio-pseuuridine, 4-methoxy-pseuuridine, 4-thio-1-methylpseuuridine, 4-thio-pseuuridine, 5-aza-uridine, dihydropseuuridine, 5-methoxyuridine, or 2′-O-methyluridine.

15. The vaccine according to claim 13, characterized in that, At least 50% of the uracil in the RNA polynucleotide is chemically modified.

16. The vaccine according to claim 15, characterized in that, At least 60% of the uracil in the RNA polynucleotide is chemically modified.

17. The vaccine according to claim 16, characterized in that, At least 70% of the uracil in the RNA polynucleotide is chemically modified.

18. The vaccine according to claim 17, characterized in that, At least 80% of the uracil in the RNA polynucleotide is chemically modified.

19. The vaccine according to claim 18, characterized in that, At least 90% of the uracil in the RNA polynucleotide is chemically modified.

20. The vaccine according to claim 19, characterized in that, The RNA polynucleotide contains 100% chemically modified uracil.

21. The vaccine according to claim 13, characterized in that, The chemical modification is located at position 5 of uracil.

22. The vaccine according to claim 11, characterized in that, The RNA polynucleotide also includes a 3'-UTR and / or at least one 5'-UTR.

23. The vaccine according to claim 22, characterized in that, The RNA polynucleotide also includes a 3'-UTR and at least one 5'-UTR.

24. The vaccine according to claim 22, characterized in that, Each of the 3'-UTR and the at least one 5'-UTR is heterogeneous to each other.

25. The vaccine according to claim 22, characterized in that, The 3'UTR is derived from genes selected from the group consisting of: housekeeping genes, genes encoding membrane proteins, genes involved in cell metabolism, genes involved in transcription, translation and replication, genes involved in protein modification, or genes involved in cell division.

26. The vaccine according to claim 22, characterized in that, The at least one 5'-UTR is derived from a gene selected from the group consisting of: housekeeping genes, genes encoding membrane proteins, genes involved in cell metabolism, genes involved in transcription, translation and replication, genes involved in protein modification, or genes involved in cell division.

27. The vaccine according to claim 11, characterized in that, The RNA polynucleotides are capped RNA polynucleotides.

28. The vaccine according to claim 27, characterized in that, The cap is selected from cap0, cap1 or cap2.

29. The vaccine according to claim 11, characterized in that, The RNA polynucleotide is a tailed RNA polynucleotide.

30. The vaccine according to claim 29, characterized in that, The tailing is performed by cleaving and polyadenylation of specific factors, cleavage stimulating factors, cleavage factor I and cleavage factor II, poly(A) polymerase, poly(A) binding protein, and / or paired proteins.

31. The vaccine according to claim 29, characterized in that, The added tail is a polyadenylated tail.

32. The vaccine according to claim 11, characterized in that, The RNA polynucleotide is a capped and tailed RNA polynucleotide.

33. The vaccine according to claim 11, characterized in that, The RNA polynucleotide consists of a 5′ cap, an open reading frame encoding the gE protein antigen variant of claim 1, and a 3′ polyadenylated tail.

34. The vaccine according to claim 33, characterized in that, The 5′ end cap is 7mG(5′)ppp(5′)N1mpNp.

35. The vaccine according to claim 33, characterized in that, The 3′ polyadenylated tail contains 20 to 300 adenine nucleotides.

36. The vaccine according to claim 35, characterized in that, The 3′ polyadenylated tail contains 40 to 200 adenine nucleotides.

37. The vaccine according to claim 36, characterized in that, The 3′ polyadenylated tail contains 50 to 100 adenine nucleotides.

38. The vaccine according to claim 37, characterized in that, The 3′ polyadenylated tail contains 60 to 70 adenine nucleotides.

39. The vaccine according to claim 10, characterized in that, The RNA vaccine is multivalent.

40. The vaccine according to claim 10, characterized in that, The RNA vaccine is formulated and delivered on a vector.

41. The vaccine according to claim 40, characterized in that, The load is selected from dendritic cells, cationic nanoemulsions, cationic peptides and polymers, liposome polymers, liposome complexes, or lipid nanoparticles.

42. The vaccine according to claim 41, characterized in that, The support medium is lipid nanoparticles.

43. The vaccine according to claim 42, characterized in that, The lipid nanoparticles are selected from cationic lipids, PEG-modified lipids, sterols and / or non-cationic lipids.

44. The vaccine according to claim 43, characterized in that, The cationic lipid is selected from the group consisting of: 2,2-dilinole-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinole-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), or 9-((4-(dimethylamino)butyryl)oxy)heptadecanedioic acid di((Z)-non-2-en-1-yl) ester (L319).

45. The vaccine according to claim 10, characterized in that, The active ingredient in the protein vaccine is the gE protein antigen variant as described in claim 1.

46. ​​The vaccine according to claim 45, characterized in that, The protein vaccine is carried by a carrier selected from serum albumin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid, diphtheria toxoid, genetically modified cross-reactive substances of diphtheria toxoid, CRM197, meningococcal outer membrane protein complex, Haemophilus influenzae protein D, rEPA, keyhole cyanin and / or flagellin.

47. The vaccine according to claim 10, characterized in that, The protein vaccine is presented by a variety of antigen presentation systems.

48. The vaccine according to claim 47, characterized in that, The antigen presentation system is selected from one or more of the following: Lys-based dendritic architecture, helper T cell epitopes, immunostimulatory lipophilic moieties, cell-penetrating peptides, free radical-induced polymerization, self-assembled nanoparticles, and gold nanoparticles.

49. The vaccine according to claim 10, characterized in that, The active ingredient in the viral vector vaccine is a nucleic acid molecule encoding an open reading frame of the gE protein antigen variant of claim 1.

50. The vaccine according to claim 49, characterized in that, The vector in the viral vector vaccine self-assembles into virus-like particles.

51. The vaccine according to claim 50, characterized in that, The carrier is a self-deactivating carrier.

52. The vaccine according to claim 51, characterized in that, The viruses are selected from lentiviruses, influenza viruses, hepatitis viruses, alpha viruses, filoviruses, adenoviruses, adeno-associated viruses, and / or flaviviruses.

53. The vaccine according to claim 52, characterized in that, The virus in question is an adenovirus or adeno-associated virus.

54. The vaccine according to claim 49, characterized in that, The nucleotide sequence is codon-optimized or codon-degenerate.

55. The vaccine according to claim 9, characterized in that, The vaccine also contains gI, gB, gH, gK, gL, gC, gN and / or gM glycoproteins of the varicella-zoster virus.

56. The vaccine according to claim 9, characterized in that, The vaccine also contains pharmaceutically acceptable adjuvants.

57. The vaccine according to claim 56, characterized in that, The pharmaceutically acceptable excipients are selected from those required for intravenous infusion, subcutaneous injection, intravenous bolus injection, intravitreal injection, or intramuscular injection.

58. A derivative of a gE protein antigen variant, characterized in that, The derivative is a complex formed by direct or indirect coupling of the gE protein antigen variant of claim 1 to a detectable marker. The detectable markers are selected from fluorescent dyes, chemiluminescent compounds, radioactive isotopes, electron-dense reagents, enzymes, colored particles, and / or biotin.

59. The use of the gE protein antigen variant of claim 1 in the preparation of a product for detecting varicella-zoster virus.

60. The use of the gE protein antigen variant of claim 1 in the preparation or screening of antibodies that recognize the varicella-zoster virus gE protein.

61. The use of the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, the cell of any one of claims 5-7, the pharmaceutical composition of claim 8, or the vaccine of any one of claims 9-57 in the preparation of a product for the prevention of disease caused by varicella-zoster virus infection.

62. The use of the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, the cell of any one of claims 5-7, the pharmaceutical composition of claim 8, or the vaccine of any one of claims 9-57 in the preparation of a product for inhibiting varicella-zoster virus.

63. The use of the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, the cell of any one of claims 5-7, the pharmaceutical composition of claim 8, or the vaccine of any one of claims 9-57 in the preparation of antibodies against gE protein.

64. The use of the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, the cell of any one of claims 5-7, or the pharmaceutical composition of claim 8 in the preparation of a varicella-zoster virus vaccine.

65. A method for screening or isolating antibodies against varicella-zoster virus gE protein, characterized in that, The method includes screening or isolating varicella-zoster virus gE protein antibodies using the gE protein antigen variant of claim 1.

66. A method for producing the gE protein antigen variant of claim 1, characterized in that, The method includes the following steps: (i) Transformed into host cells using the vector according to claim 3 or 4, or directly using cells according to any one of claims 5-7; (ii) Culturing the host cells or cells from step (i) under suitable conditions; (iii) The gE protein antigen variant of claim 1 is isolated and purified from the host cell culture medium.

67. A method for preparing antibodies against varicella-zoster virus gE protein, characterized in that, Antibodies were obtained by immunizing non-human animals using the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, or the vector of claim 3 or 4.

68. The method according to claim 67, characterized in that, The non-human animals mentioned are vertebrates.

69. The method according to claim 68, characterized in that, The vertebrates are selected from mice, rats, guinea pigs, rabbits, sheep, or non-human primates.

70. A detection / prevention reagent, test strip, and kit comprising the same, characterized in that, The vaccine comprises the gE protein antigen variant of claim 1, the nucleic acid molecule of claim 2, the vector of claim 3 or 4, the cell of any one of claims 5-7, the pharmaceutical composition of claim 8, the vaccine of any one of claims 9-57, or the derivative of claim 58.

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