Novel cell membrane targeting signal peptides and uses thereof
By designing novel signal peptides, the problems of post-translational control and targeted localization of encoded proteins within cells were solved, achieving efficient targeted localization and delivery of encoded proteins and improving the therapeutic efficacy of therapeutic mRNAs.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- BOARD OF RGT THE UNIV OF TEXAS SYST
- Filing Date
- 2024-10-09
- Publication Date
- 2026-07-14
AI Technical Summary
Existing technologies lack effective means for post-translational control and targeted localization of encoded proteins, resulting in low efficiency in protein delivery and localization of therapeutic mRNAs.
A series of novel signal peptides, containing specific amino acid sequences, have been designed to guide the post-translational localization and targeting of encoded proteins to the cell membrane. These include combinations and variants of various signal peptides that can form recombinant polypeptides with encoded proteins to achieve precise intracellular transport and secretion.
It improves the targeting efficiency and delivery effect of encoded proteins, enhances the therapeutic potential of therapeutic mRNAs, and meets the targeting and secretion needs of encoded proteins in vivo.
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Abstract
Description
Related applications
[0001] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 588,916, filed October 9, 2023, and U.S. Provisional Application Serial No. 63 / 605,387, filed December 1, 2023, each of which is incorporated herein by reference in its entirety.
[0002] sequence list This application contains references to amino acid sequences and / or nucleic acid sequences, which were submitted with this application as a sequence listing XML file named "825591_sequencelisting.xml", with a size of 36,864 bytes, created on October 9, 2024. The above sequence listing is incorporated herein by reference in its entirety in accordance with 37 CFR §1.52(e)(5). Technical Field
[0003] This disclosure relates generally to novel signal peptides and more specifically to novel signal peptides for targeting encoded proteins to the cell membrane. Background Technology
[0004] Recent scientific discoveries have highlighted the numerous therapeutic applications of mRNA due to its modular nature and ability to provide customizable "instructions" to produce functional proteins. Clinical studies continue to confirm its beneficial efficacy, further emphasizing its potential as a next-generation genetic drug, where scientists are only beginning to scratch the surface of its potential applications. To date, most research has focused on enhancing the intracellular expression of encoded peptides. However, little attention has been paid to the post-translational control of the encoded protein and its targeted localization and / or secretion in vivo.
[0005] Within the cell, various unique pathways and processes exist that allow proteins to shuttle to various organelles and to be exported to the extracellular space via excretion pathways. However, in order to utilize these transport systems, the mRNA encoding each protein must also contain a metaphorical transport marker called a signal peptide (SP) upstream of the protein sequence. Designing novel SPs for therapeutic mRNAs can greatly benefit the delivery, localization, and clearance of the encoded therapeutic proteins. This disclosure addresses these and other needs. Summary of the Invention
[0006] In some embodiments, a signal peptide is provided. In some embodiments, the signal peptide comprises an amino acid sequence selected from the group consisting of formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII, XXXIV, XXXV, XXXVI, XXXVII, XXXVIII, or XXXIX; Where equation I is expressed as: A1-A2-A3-A4-A5-A6-A7-A8-A9-A 10 -A 11 -A 12 -A 13 -A 14 (Formula I) And A1-A 14 Their identities are provided in Table 2; Equation II is expressed as: B1-B2-B3-B4-B5-B6-B7-B8-B9-B 10 -B 11 -B 12 -B 13 -B 14 -B 15 (Formula II) And B1-B 15 Their identities are provided in Table 3; Equation III is expressed as: C1-C2-C3-C4-C5-C6-C7-C8-C9-C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 (Formula III) And C1-C 16 Their identities are provided in Table 4; Where equation IV is represented as: D1-D2-D3-D4-D5-D6-D7-D8-D9-D 10 -D 11 -D 12 -D 13 -D 14 -D 15 -D 16 -D 17(Formula IV) And D1-D 17 Their identities are provided in Table 5; Where V is expressed as: E1-E2-E3-E4-E5-E6-E7-E8-E9-E 10 -E 11 -E 12 -E 13 -E 14 -E 15 -E 16 -E 17 -E 18 (Formula V) And E1-E 18 Their identities are provided in Table 6; Wherein, VI is represented as: F1-F2-F3-F4-F5-F6-F7-F8-F9-F 10 -F 11 -F 12 -F 13 -F 14 -F 15 -F 16 -F 17 -F 18 -F 19 (Form VI) And F1-F 19 Their identities are provided in Table 7; Equation VII is expressed as: G1-G2-G3-G4-G5-G6-G7-G8-G9-G 10 -G 11 -G 12 -G 13 -G 14 -G 15 -G 16 -G 17 -G 18 -G 19 -G 20 (Equation VII) And G1-G 20 Their identities are provided in Table 8; Where equation VIII is expressed as: H1-H2-H3-H4-H5-H6-H7-H8-H9-H 10 -H 11 -H 12 -H 13 -H 14 -H 15 -H 16 -H17 -H 18 -H 19 -H 20 -H 21 (Formula VIII) And H1-H 21 Their identities are provided in Table 9; Where IX is represented as: I1-I2-I3-I4-I5-I6-I7-I8-I9-I 10 -I 11 -I 12 -I 13 -I 14 -I 15 -I 16 -I 17 -I 18 -I 19 -I 20 -I 21 -I 22 (Form IX) And I1-I 22 Their identities are provided in Table 10; Where X is represented as: J1-J2-J3-J4-J5-J6-J7-J8-J9-J 10 -J 11 -J 12 -J 13 -J 14 -J 15 -J 16 -J 17 -J 18 -J 19 -J 20 -J 21 -J 22 -J 23 (Formula X) And J1-J 23 Their identities are provided in Table 11; Where XI is represented as: K1-K2-K3-K4-K5-K6-K7-K8-K9-K 10 -K 11 -K 12 -K 13 -K 14 -K 15 -K 16 -K 17 -K 18 -K 19 -K 20 -K 21 -K 22-K 23 -K 24 (Formula XI) And K1-K 24 Their identities are provided in Table 12; Where XII is expressed as: L1-L2-L3-L4-L5-L6-L7-L8-L9-L 10 -L 11 -L 12 -L 13 -L 14 -L 15 -L 16 -L 17 -L 18 -L 19 -L 20 -L 21 -L 22 -L 23 -L 24 -L 25 (Formula XII) And L1-L 25 Their identities are provided in Table 13; Where XIII is expressed as: M1-M2-M3-M4-M5-M6-M7-M8-M9-M 10 -M 11 -M 12 -M 13 -M 14 -M 15 -M 16 -M 17 -M 18 -M 19 -M 20 -M 21 -M 22 -M 23 -M 24 -M 25 -M 26 (Formula XIII) And M1-M 26 Their identities are provided in Table 14; Where XIV is expressed as: N1-N2-N3-N4-N5-N6-N7-N8-N9-N 10 -N 11 -N 12 -N 13 -N 14 -N 15 -N 16 -N 17 -N 18 -N19 -N 20 -N 21 -N 22 -N 23 -N 24 -N 25 -N 26 -N 27 (Form XIV) And N1-N 27 Their identities are provided in Table 15; Where XV is expressed as: O1-O2-O3-O4-O5-O6-O7-O8-O9-O 10 -O 11 -O 12 -O 13 -O 14 -O 15 -O 16 -O 17 -O 18 -O 19 -O 20 -O 21 -O 22 -O 23 -O 24 -O 25 -O 26 -O 27 -O 28 (Formula XV) And O1-O 28 Their identities are provided in Table 16; Where XVI is expressed as: P1-P2-P3-P4-P5-P6-P7-P8-P9-P 10 -P 11 -P 12 -P 13 -P 14 -P 15 -P 16 -P 17 -P 18 -P 19 -P 20 -P 21 -P 22 -P 23 -P 24 -P 25 -P 26 -P 27 -P 28 -P 29 (Formula XVI) And P1-P 29 Their identities are provided in Table 17; Where XVII is expressed as: Q1-Q2-Q3-Q4-Q5-Q6-Q7-Q8-Q9-Q 10 -Q 11 -Q 12 -Q 13 -Q 14 -Q 15 -Q 16 -Q 17 -Q 18 -Q 19 -Q 20 -Q 21 -Q 22 -Q 23 -Q 24 -Q 25 -Q 26 -Q 27 -Q 28 -Q 29 -Q 30 (Form XVII) And Q1-Q 30 Their identities are provided in Table 18; Where XVIII is expressed as: R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 (Form XVIII) And R1-R 31 Their identities are provided in Table 19; Where XIX is represented as: S1-S2-S3-S4-S5-S6-S7-S8-S9-S 10 -S 11 -S 12 -S 13 -S 14 -S15 -S 16 -S 17 -S 18 -S 19 -S 20 -S 21 -S 22 -S 23 -S 24 -S 25 -S 26 -S 27 -S 28 -S 29 -S 30 -S 31 -S 32 (Form XIX) And S1-S 32 The identities are provided in Table 20; Where XX is represented as: T1-T2-T3-T4-T5-T6-T7-T8-T9-T 10 -T 11 -T 12 -T 13 -T 14 -T 15 -T 16 -T 17 -T 18 -T 19 -T 20 -T 21 -T 22 -T 23 -T 24 -T 25 -T 26 -T 27 -T 28 -T 29 -T 30 -T 31 -T 32 -T 33 (Formula XX) And T1-T 33 Their identities are provided in Table 21; Where XXI is represented as: U1-U2-U3-U4-U5-U6-U7-U8-U9-U 10 -U 11 -U 12 -U 13 -U 14 -U 15 -U 16 -U 17 -U 18 -U 19 -U20 -U 21 -U 22 -U 23 -U 24 -U 25 -U 26 -U 27 -U 28 -U 29 -U 30 -U 31 -U 32 -U 33 -U 34 (Formula XXI) And U1-U 34 Their identities are provided in Table 22; Where equation XXII is represented as: V1-V2-V3-V4-V5-V6-V7-V8-V9-V 10 -V 11 -V 12 -V 13 -V 14 -V 15 -V 16 -V 17 -V 18 -V 19 -V 20 -V 21 -V 22 -V 23 -V 24 -V 25 -V 26 -V 27 -V 28 -V 29 -V 30 -V 31 -V 32 -V 33 -V 34 -V 35 (Formula XXII) And V1-V 35 Their identities are provided in Table 23; Where equation XXIII is represented as: W1-W2-W3-W4-W5-W6-W7-W8-W9-W 10 -W 11 -W 12 -W 13 -W 14 -W 15 -W 16 -W 17 -W 18 -W 19 -W20 -W 21 -W 22 -W 23 -W 24 -W 25 -W 26 -W 27 -W 28 -W 29 -W 30 -W 31 -W 32 -W 33 -W 34 -W 35 -W 36 (Formula XXIII) And W1-W 36 Their identities are provided in Table 24; Where equation XXIV is represented as: X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36 -X 37 (Formula XXIV) And X1-X 37 The identities are provided in Table 25; Where XXV is represented as: Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y 10 -Y 11 -Y 12 -Y 13 -Y 14 -Y 15 -Y16 -Y 17 -Y 18 -Y 19 -Y 20 -Y 21 -Y 22 -Y 23 -Y 24 -Y 25 -Y 26 -Y 27 -Y 28 -Y 29 -Y 30 -Y 31 -Y 32 -Y 33 -Y 34 -Y 35 -Y 36 -Y 37 -Y 38 (Formula XXV) And Y1-Y 38 Their identities are provided in Table 26; Wherein, XXVI is represented as: Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -Z 20 -Z 21 -Z 22 -Z 23 -Z 24 -Z 25 -Z 26 -Z 27 -Z 28 -Z 29 -Z 30 -Z 31 -Z 32 -Z 33 -Z 34 -Z 35 -Z 36 -Z 37 -Z 38 -Z 39 (Form XXVI) And Z1-Z 39 Their identities are provided in Table 27; Where equation XXVII is represented as: (AA)1-(AA)2-(AA)3-(AA)4-(AA)5-(AA)6-(AA)7-(AA)8-(AA)9-(AA) 10 -(AA) 11 -(AA) 12 -(AA) 13 -(AA) 14 -(AA) 15 -(AA) 16 -(AA) 17 -(AA) 18 -(AA) 19 -(AA) 20 -(AA) 21 -(AA) 22 -(AA) 23 -(AA) 24 -(AA) 25 -(AA) 26 -(AA) 27 -(AA) 28 -(AA) 29 23 -(BB) 24 -(BB) 25 -(BB) 26 -(BB) 27 -(BB) 28 -(BB) 29 -(BB) 30 -(BB) 31 -(BB) 32 -(BB) 33 -(BB) 34 -(BB) 35 -(BB) 36 -(BB) 37 -(BB) 38 -(BB) 39 -(BB) 40 -(BB) 41 (Formula XXVIII) And (BB)1-(BB) 41 Their identities are provided in Table 29; Where XXIX is represented as: (CC)1-(CC)2-(CC)3-(CC)4-(CC)5-(CC)6-(CC)7-(CC)8-(CC)9-(CC) 10 -(CC) 11 -(CC) 12 -(CC) 13 -(CC) 14 -(CC) 15 -(CC) 16 -(CC) 17 -(CC) 18 -(CC) 19 -(CC) 20 -(CC) 21 -(CC) 22 -(CC) 23 -(CC) 24 -(CC) 25 -(CC) 26 -(CC) 27 -(CC) 28 -(CC) 29 -(CC) 30 -(CC) 31 -(CC) 32 -(CC) 33 -(CC) 34 -(CC) 35 -(CC) 36 -(CC) 37 -(CC)38 -(CC) 39 -(CC) 40 -(CC) 41 -(CC) 42 (Formula XXIX) And (CC)1-(CC) 42 Their identities are provided in Table 30; Where the formula XXX is represented as: (DD)1-(DD)2-(DD)3-(DD)4-(DD)5-(DD)6-(DD)7-(DD)8-(DD)9-(DD) 10 -(DD) 11 -(DD) 12 -(DD) 13 -(DD) 14 -(DD) 15 -(DD) 16 -(DD) 17 -(DD) 18 -(DD) 19 -(DD) 20 -(DD) 21 -(DD) 22 -(DD) 23 -(DD) 24 -(DD) 25 -(DD) 26 -(DD) 27 -(DD) 28 -(DD) 29 -(DD) 30 -(DD) 31 -(DD) 32 -(DD) 33 -(DD) 34 -(DD) 35 -(DD) 36 -(DD) 37 -(DD) 38 -(DD) 39 -(DD) 40 -(DD) 41 -(DD) 42 -(DD) 43 (Formula XXX) And (DD)1-(DD) 43 Their identities are provided in Table 31; Wherein, XXXI is represented as: (EE)1-(EE)2-(EE)3-(EE)4-(EE)5-(EE)6-(EE)7-(EE)8-(EE)9-(EE)10 -(EE) 11 -(EE) 12 -(EE) 13 -(EE) 14 -(EE) 15 -(EE) 16 -(EE) 17 -(EE) 18 -(EE) 19 -(EE) 20 -(EE) 21 -(EE) 22 -(EE) 23 -(EE) 24 -(EE) 25 -(EE) 26 -(EE) 27 -(EE) 28 -(EE) 29 -(EE) 30 -(EE) 31 -(EE) 32 -(EE) 33 -(EE) 34 -(EE) 35 -(EE) 36 -(EE) 37 -(EE) 38 -(EE) 39 -(EE) 40 -(EE) 41 -(EE) 42 -(EE) 43 -(EE) 44 (Formula XXXI) And (EE)1-(EE) 44 Their identities are provided in Table 32; Wherein, equation XXXII is represented as: (FF)1-(FF)2-(FF)3-(FF)4-(FF)5-(FF)6-(FF)7-(FF)8-(FF)9-(FF) 10 -(FF) 11 -(FF) 12 -(FF) 13 -(FF) 14 -(FF) 15 -(FF) 16 -(FF) 17 -(FF) 18 -(FF) 19 -(FF) 20 -(FF) 21 -(FF)22 -(FF) 23 -(FF) 24 -(FF) 25 -(FF) 26 -(FF) 27 -(FF) 28 -(FF) 29 -(FF) 30 -(FF)33 -(GG) 34 -(GG) 35 -(GG) 36 -(GG) 37 -(GG) 38 -(GG) 39 -(GG) 40 -(GG) 41 -(GG) 42 -(GG) 43 -(GG)<000D723>-(GG) 45 -(GG) 46 (Formula XXXIII) and (GG)1-(GG) 46 is as provided in Table 34; where Formula XXXIV is represented as: (HH)1-(HH)2-(HH)I3-(HH)4-(HH)5-(HH)6-(HH)7-(HH)8-(HH)9-(HH) 10 -(HH) 11 -(HH) 12 -(HH) 13 -(HH) 14 -(HH) 15 -(HH) 16 -(HH) 17 -(HH) 18 -(HH) 19 -(HH) 20 -(HH) 21 -(HH) 22 -(HH) 23 -(HH) 24 -(HH) 25 -(HH) 26 -(HH) 27 -(HH) 28 -(HH) 29 -(HH) 30 -(HH) 31 -(HH) 32 -(HH) 33 -(HH) 34 -(HH) 35 -(HH) 36 -(HH) 37 -(HH) 38 -(HH) 39 -(HH) 40 -(HH) 41 -(HH) 42 -(HH)43 -(HH) 44 -(HH) 45 -(HH) 46 -(HH) 47 (Formula XXXIV) And (HH)1-(HH) 47 Their identities are provided in Table 35; Wherein, XXXV is represented as: (II)1-(II)2-(II)3-(II)4-(II)5-(II)6-(II)7-(II)8-(II)9-(II) 10 -(II) 11 -(II) 12 -(II) 13 -(II) 14 -(II) 15 -(II) 16 -(II) 17 -(II) 18 -(II) 19 -(II) 20 -(II) 21 -(II) 22 -(II) 23 -(II) 24 -(II) 25 -(II) 26 -(II) 27 -(II) 28 -(II) 29 -(II) 30 -(II) 31 -(II) 32 -(II) 33 -(II) 34 -(II) 35 -(II) 36 -(II) 37 -(II) 38 -(II) 39 -(II) 40 -(II) 41 -(II) 42 -(II) 43 -(II) 44 -(II) 45 -(II) 46 -(II) 47 -(II) 48 (Formula XXXV) And (II)1-(II) 48 The identities are provided in Table 36; Wherein, XXXVI is represented as: (JJ)1-(JJ)2-(JJ)3-(JJ)4-(JJ)5-(JJ)6-(JJ)7-(JJ)8-(JJ)9-(JJ) 10 -(JJ) 11 -(JJ) 12 -(JJ) 13 -(JJ) 14 -(JJ) 15 -(JJ) 16 -(JJ) 17 -(JJ) 18 -(JJ) 19 -(JJ) 20 -(JJ) 21 -(JJ) 22 -(JJ) 23 -(JJ) 24 -(JJ) 25 -(JJ) 26 -(JJ) 27 -(JJ) 28 -(JJ) 29 -(JJ) 30 -(JJ) 31 -(JJ) 32 -(JJ) 33 -(JJ) 34 -(JJ) 35 -(JJ) 36 -(JJ) 37 -(JJ) 38 -(JJ) 39 -(JJ) 40 -(JJ) 41 -(JJ) 42 -(JJ) 43 -(JJ) 44 -(JJ) 45 -(JJ) 46 -(JJ) 47 -(JJ) 48 -(JJ) 49 (Formula XXXVI) And (JJ)1-(JJ) 49 Their identities are provided in Table 37; Where equation XXXVII is represented as: (KK)1-(KK)2-(KK)3-(KK)4-(KK)5-(KK)6-(KK)7-(KK)8-(KK)9-(KK) 10 -(KK) 11 -(KK) 12-(KK) 13 -(KK) 14 -(KK) 15 -(KK) 16 -(KK) 17 -(KK) 18 -(KK) 19 -(KK) 20 -(KK) 21 -(KK) 22 -(KK) 23 -(KK) 24 -(KK) 25 -(KK) 26 -(KK) 27 -(KK) 28 -(KK) 29 -(KK) 30 -(KK) 31 -(KK) 32 -(KK) 33 -(KK) 34 -(KK) 35 -(KK) 36 -(KK) 37 -(KK) 38 -(KK) 39 -(KK) 40 -(KK) 41 -(KK) 42 -(KK) 43 -(KK) 44 -(KK) 45 -(KK) 46 -(KK) 47 -(KK) 48 -(KK) 49 -(KK) 50 -(KK) 51 -(KK) 52 (Formula XXXVII) And (KK)1-(KK) 52 The identities are provided in Table 38; Wherein, equation XXXVIII is represented as: (LL)1-(LL)2-(LL)3-(LL)4-(LL)5-(LL)6-(LL)7-(LL)8-(LL)9-(LL) 10 -(LL) 11 -(LL) 12 -(LL) 13 -(LL) 14 -(LL) 15 -(LL) 16-(LL) 17 -(LL) 18 -(LL) 19 -(LL) 20 -(LL) 21 -(LL) 22 -(LL) 23 -(LL) 24 -(LL) 25 -(LL) 26 -(LL) 27 -(LL) 28 -(LL) 29 -(LL) 30 -(LL) 31 -(LL) 32 -(LL) 33 -(LL) 34 -(LL) 35 -(LL) 36 -(LL) 37 -(LL) 38 -(LL) 39 -(LL) 40 -(LL) 41 -(LL) 42 -(LL) 43 -(LL) 44 -(LL) 45 -(LL) 46 -(LL) 47 -(LL) 48 -(LL) 49 -(LL) 50 -(LL) 51 -(LL) 52 -(LL) 53 -(LL) 54 -(LL) 55 (Formula XXXVIII) and (LL)1-(LL) 55 has the identity as provided in Table 39; and where Formula XXXIX is represented as: (MM)1-(MM)2-(MM)3-(MM)4-(MM)5-(MM)6-(MM)7-(MM)8-(MM)9-(MM) 10 -(MM) 11 -(MM) 12 -(MM) 13 -(MM) 14 -(MM) 15 -(MM) 16 -(MM) 17-(MM) 18 -(MM) 19 -(MM) 20 -(MM) 21 -(MM) 22 -(MM) 23 -(MM) 24 -(MM) 25 -(MM) 26 -(MM) 27 -(MM) 28 -(MM) 29 -(MM) 30 -(MM) 31 -(MM) 32 -(MM) 33 -(MM) 34 -(MM) 35 -(MM) 36 -(MM) 37 -(MM) 38 -(MM) 39 -(MM) 40 -(MM) 41 -(MM) 42 -(MM) 43 -(MM)<M 44 -(MM) 45 -(MM) 46 -(MM) 47 -(MM) 48 -(MM) 49 -(MM) 50 -(MM) 51 -(MM) 52 -(MM) 53 -(MM) 54 -(MM) 55 -(MM) 56 -(MM) 57 -(MM) 58 -(MM) 59 (Formula XXXIX) and (MM)1-(MM) 59 The identity of is provided in Table 40.
[0007] In some embodiments, the signal peptide comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 9, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
[0008] In some embodiments, a signal peptide is provided comprising an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 9, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
[0009] In some implementations, the signal peptides provided herein are novel signal peptides.
[0010] In some embodiments, a recombinant polypeptide is provided. In some embodiments, the recombinant polypeptide comprises X1-(Y1). a -Z1, where X1 is the signal peptide, Y1 is the peptide linker, and Z1 is the payload protein, where a is an integer selected from 0 and 1.
[0011] In some embodiments, signal peptide X1 comprises an amino acid sequence selected from the group consisting of formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII, XXXIV, XXXV, XXXVI, XXXVII, XXXVIII, or XXXIX; Where equation I is expressed as: A1-A2-A3-A4-A5-A6-A7-A8-A9-A 10 -A11 -A 12 -A 13 -A 14 (Formula I) And A1-A 14 Their identities are provided in Table 2; Equation II is expressed as: B1-B2-B3-B4-B5-B6-B7-B8-B9-B 10 -B 11 -B 12 -B 13 -B 14 -B 15 (Formula II) And B1-B 15 Their identities are provided in Table 3; Equation III is expressed as: C1-C2-C3-C4-C5-C6-C7-C8-C9-C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 (Formula III) And C1-C 16 Their identities are provided in Table 4; Where equation IV is represented as: D1-D2-D3-D4-D5-D6-D7-D8-D9-D 10 -D 11 -D 12 -D 13 -D 14 -D 15 -D 16 -D 17 (Formula IV) And D1-D 17 Their identities are provided in Table 5; Where V is expressed as: E1-E2-E3-E4-E5-E6-E7-E8-E9-E 10 -E 11 -E 12 -E 13 -E 14 -E 15 -E 16 -E 17 -E 18 (Formula V) And E1-E 18 Their identities are provided in Table 6; Wherein, VI is represented as: F1-F2-F3-F4-F5-F6-F7-F8-F9-F 10 -F 11 -F 12 -F 13 -F 14 -F 15 -F 16 -F 17 -F 18 -F 19 (Form VI) And F1-F 19 Their identities are provided in Table 7; Equation VII is expressed as: G1-G2-G3-G4-G5-G6-G7-G8-G9-G 10 -G 11 -G 12 -G 13 -G 14 -G 15 -G 16 -G 17 -G 18 -G 19 -G 20 (Equation VII) And G1-G 20 Their identities are provided in Table 8; Where equation VIII is expressed as: H1-H2-H3-H4-H5-H6-H7-H8-H9-H 10 -H 11 -H 12 -H 13 -H 14 -H 15 -H 16 -H 17 -H 18 -H 19 -H 20 -H 21 (Formula VIII) And H1-H 21 Their identities are provided in Table 9; Where IX is represented as: I1-I2-I3-I4-I5-I6-I7-I8-I9-I 10 -I 11 -I 12 -I 13 -I 14 -I 15 -I 16 -I 17 -I18 -I 19 -I 20 -I 21 -I 22 (Form IX) And I1-I 22 Their identities are provided in Table 10; Where X is represented as: J1-J2-J3-J4-J5-J6-J7-J8-J9-J 10 -J 11 -J 12 -J 13 -J 14 -J 15 -J 16 -J 17 -J 18 -J 19 -J 20 -J 21 -J 22 -J 23 (Formula X) And J1-J 23 Their identities are provided in Table 11; Where XI is represented as: K1-K2-K3-K4-K5-K6-K7-K8-K9-K 10 -K 11 -K 12 -K 13 -K 14 -K 15 -K 16 -K 17 -K 18 -K 19 -K 20 -K 21 -K 22 -K 23 -K 24 (Formula XI) And K1-K 24 Their identities are provided in Table 12; Where XII is expressed as: L1-L2-L3-L4-L5-L6-L7-L8-L9-L 10 -L 11 -L 12 -L 13 -L 14 -L 15 -L 16 -L 17 -L 18 -L 19 -L 20-L 21 -L 22 -L 23 -L 24 -L 25 (Formula XII) And L1-L 25 Their identities are provided in Table 13; Where XIII is expressed as: M1-M2-M3-M4-M5-M6-M7-M8-M9-M 10 -M 11 -M 12 -M 13 -M 14 -M 15 -M 16 -M 17 -M 18 -M 19 -M 20 -M 21 -M 22 -M 23 -M 24 -M 25 -M 26 (Formula XIII) And M1-M 26 Their identities are provided in Table 14; Where XIV is expressed as: N1-N2-N3-N4-N5-N6-N7-N8-N9-N 10 -N 11 -N 12 -N 13 -N 14 -N 15 -N 16 -N 17 -N 18 -N 19 -N 20 -N 21 -N 22 -N 23 -N 24 -N 25 -N 26 -N 27 (Form XIV) And N1-N 27 Their identities are provided in Table 15; Where XV is expressed as: O1-O2-O3-O4-O5-O6-O7-O8-O9-O 10 -O 11 -O 12 -O 13 -O14 -O 15 -O 16 -O 17 -O 18 -O 19 -O 20 -O 21 -O 22 -O 23 -O 24 -O 25 -O 26 -O 27 -O 28 (Formula XV) And O1-O 28 Their identities are provided in Table 16; Where XVI is expressed as: P1-P2-P3-P4-P5-P6-P7-P8-P9-P 10 -P 11 -P 12 -P 13 -P 14 -P 15 -P 16 -P 17 -P 18 -P 19 -P 20 -P 21 -P 22 -P 23 -P 24 -P 25 -P 26 -P 27 -P 28 -P 29 (Formula XVI) And P1-P 29 Their identities are provided in Table 17; Where XVII is expressed as: Q1-Q2-Q3-Q4-Q5-Q6-Q7-Q8-Q9-Q 10 -Q 11 -Q 12 -Q 13 -Q 14 -Q 15 -Q 16 -Q 17 -Q 18 -Q 19 -Q 20 -Q 21 -Q 22 -Q 23 -Q 24 -Q 25 -Q 26-Q 27 -Q 28 -Q 29 -Q 30 (Form XVII) And Q1-Q 30 Their identities are provided in Table 18; Where XVIII is expressed as: R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 (Form XVIII) And R1-R 31 Their identities are provided in Table 19; Where XIX is represented as: S1-S2-S3-S4-S5-S6-S7-S8-S9-S 10 -S 11 -S 12 -S 13 -S 14 -S 15 -S 16 -S 17 -S 18 -S 19 -S 20 -S 21 -S 22 -S 23 -S 24 -S 25 -S 26 -S 27 -S 28 -S 29 -S 30 -S 31 -S 32 (Form XIX) And S1-S 32The identities are provided in Table 20; Where XX is represented as: T1-T2-T3-T4-T5-T6-T7-T8-T9-T 10 -T 11 -T 12 -T 13 -T 14 -T 15 -T 16 -T 17 -T 18 -T 19 -T 20 -T 21 -T 22 -T 23 -T 24 -T 25 -T 26 -T 27 -T 28 -T 29 -T 30 -T 31 -T 32 -T 33 (Formula XX) And T1-T 33 Their identities are provided in Table 21; Where XXI is represented as: U1-U2-U3-U4-U5-U6-U7-U8-U9-U 10 -U 11 -U 12 -U 13 -U 14 -U 15 -U 16 -U 17 -U 18 -U 19 -U 20 -U 21 -U 22 -U 23 -U 24 -U 25 -U 26 -U 27 -U 28 -U 29 -U 30 -U 31 -U 32 -U 33 -U 34 (Formula XXI) And U1-U 34 Their identities are provided in Table 22; Where equation XXII is represented as: V1-V2-V3-V4-V5-V6-V7-V8-V9-V 10 -V 11 -V 12 -V 13 -V 14 -V 15 -V 16 -V 17 -V 18 -V 19 -V 20 -V 21 -V 22 -V 23 -V 24 -V 25 -V 26 -V 27 -V 28 -V 29 -V 30 -V 31 -V 32 -V 33 -V 34 -V 35 (Formula XXII) And V1-V 35 Their identities are provided in Table 23; Where equation XXIII is represented as: W1-W2-W3-W4-W5-W6-W7-W8-W9-W 10 -W 11 -W 12 -W 13 -W 14 -W 15 -W 16 -W 17 -W 18 -W 19 -W 20 -W 21 -W 22 -W 23 -W 24 -W 25 -W 26 -W 27 -W 28 -W 29 -W 30 -W 31 -W 32 -W 33 -W 34 -W 35 -W 36 (Formula XXIII) And W1-W 36 Their identities are provided in Table 24; Where equation XXIV is represented as: X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36 -X 37 (Formula XXIV) And X1-X 37 The identities are provided in Table 25; Where XXV is represented as: Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y 10 -Y 11 -Y 12 -Y 13 -Y 14 -Y 15 -Y 16 -Y 17 -Y 18 -Y 19 -Y 20 -Y 21 -Y 22 -Y 23 -Y 24 -Y 25 -Y 26 -Y 27 -Y 28 -Y 29 -Y 30 -Y 31 -Y 32 -Y 33 -Y 34 -Y 35 -Y 36 -Y37 -Y 38 (Formula XXV) And Y1-Y 38 Their identities are provided in Table 26; Wherein, XXVI is represented as: Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -Z 20 -Z 21 -Z 22 -Z 23 -Z 24 -Z 25 -Z 26 -Z 27 -Z 28 -Z 29 -Z 30 -Z 31 -Z 32 -Z 33 -Z 34 -Z 35 -Z 36 -Z 37 -Z 38 -Z 39 (Form XXVI) And Z1-Z 39 Their identities are provided in Table 27; Where equation XXVII is represented as: (AA)1-(AA)2-(AA)3-(AA)4-(AA)5-(AA)6-(AA)7-(AA)8-(AA)9-(AA) 10 -(AA) 11 -(AA) 12 -(AA) 13 -(AA) 14 -(AA) 15 -(AA) 16 -(AA) 17 -(AA) 18 -(AA) 19 -(AA) 20 -(AA) 21 -(AA) 22 -(AA) 23 -(AA) 24-(AA) 25 -(AA) 26 -(AA) 27 -(AA) 28 -(AA) 29 -(AA) 30 -(AA) 31 -(AA) 32 -(AA) 33 -(AA) 34 -(AA) 35 -(AA) 36 -(AA) 37 -(AA) 38 -(AA) 39 -(AA) 40 (Formula XXVII) and (AA)1-(AA) 40 has the identity as provided in Table 28; where Formula XXVIII is represented as: (BB)1-(BB)2-(BB)3-(BB)4-(BB)5-(BB)6-(BB)7-(BB)8-(BB)9-(BB) 10 -(BB) 11 -(BB) 12 -(BB) 13 -(BB) 14 -(BB) 15 -(BB)<0041 (Formula XXVIII) And (BB)1-(BB) 41 Their identities are provided in Table 29; Where XXIX is represented as: (CC)1-(CC)2-(CC)3-(CC)4-(CC)5-(CC)6-(CC)7-(CC)8-(CC)9-(CC) 10 -(CC) 11 -(CC) 12 -(CC) 13 -(CC) 14 -(CC) 15 -(CC) 16 -(CC) 17 -(CC) 18 -(CC) 19 -(CC) 20 -(CC) 21 -(CC) 22 -(CC) 23 -(CC) 24 -(CC) 25 -(CC) 26 -(CC) 27 -(CC) 28 -(CC) 29 -(CC) 30 -(CC) 31 -(CC) 32 -(CC) 33 -(CC) 34 -(CC) 35 -(CC) 36 -(CC) 37 -(CC) 38 -(CC) 39 -(CC) 40 -(CC) 41 -(CC) 42 (Formula XXIX) And (CC)1-(CC) 42 Their identities are provided in Table 30; Where the formula XXX is represented as: (DD)1-(DD)2-(DD)3-(DD)4-(DD)5-(DD)6-(DD)7-(DD)8-(DD)9-(DD) 10 -(DD) 11 -(DD) 12 -(DD) 13 -(DD) 14 -(DD)15 -(DD) 16 -(DD) 17 -(DD) 18 -(DD) 19 -(DD) 20 -(DD) 21 -(DD) 22 -(DD) 23 -(DD) 24 -(DD) 25 -(DD) 26 -(DD) 27 -(DD) 28 -(DD) 29 -(DD) 30 -(DD) 31 -(DD) 32 -(DD) 33 -(DD) 34 -(DD) 35 -(DD) 36 -(DD) 37 -(DD) 38 -(DD) 39 -(DD) 40 -(DD) 41 -(DD) 42 -(DD) 43 (Formula XXX) and (DD)1-(DD) 43 The identity of... is provided in Table 31; where Formula XXXI is represented as: (EE)1-(EE)2-(EE)3-(EE)4-(EE)5-(EE)6-(EE)7-(EE)8-(EE)9-(EE) 10 -(EE) 11 -(EE) 12 -(EE) 13 -(EE) 14 -(EE)[[ID=七十六]] 15 [[ID=七十七]]-(EE)[[ID=七十八]] 16 [[ID=七十九]]-(EE)[[ID=八十]] 17 [[ID=八十一]]-(EE)[[ID=八十二]] 18 [[ID=八十三]]-(EE)[[ID=八十四]] 19 [[ID=八十五]]-(EE)[[ID=八十六]] 20 [[ID=八十七]]-(EE)[[ID=八十八]] 21 [[ID=八十九]]-(EE)[[ID=九十]] 22 [[ID=九十一]]-(EE)[[ID=九十二]] 23 [[ID=九十三]]-(EE)[[ID=九十四]] 24 [[ID=九十五]]-(EE)[[ID=九十六]] 25 [[ID=九十七]]-(EE)[[ID=九十八]] 26 [[ID=九十九]]-(EE)[[ID=一百]] 27 [[ID=一百零一]]-(EE)[[ID=一百零二]] 28 请注意,原文中“ 15 ”到“ 17 ”之间的翻译内容中,原内容里的“”等编号在翻译中被直接替换为了对应的中文编号表述,可能会与原文的编号标识方式不一致,这是为了完全按照您提供的规则进行翻译。如果有特殊要求,请进一步说明。-(EE) 29 -(EE) 30 -(EE) 31 -(EE) 32 -(EE) 33 -(EE) 34 -(EE) 35 -(EE) 36 -(EE) 37 -(EE) 38 -(EE) 39 -(EE) 40 -(EE) 41 -(EE) 42 -(EE) 43 -(EE) 44 (Formula XXXI) and (EE)1-(EE) 44 has the identity as provided in Table 32; where Formula XXXII is represented as: (FF)1-(FF)2-(FF)3-(FF)L-(FF)5-(FF)6-(FF)7-(FF)8-(FF)9-(FF) 10 -(FF) 11 -(FF) 12 -(FF) 13 -(FF) 14 -(FF) 15 -(FF) 16 -(FF) 17 -(FF) 18 -(FF) 19 -(FF) 20 -(FF) 21 -(FF) 22 -(FF) 23 -(FF) 24 -(FF) 25 -(FF) 26 -(FF) 27 -(FF) 28 -(FF) 29 -(FF) 30 -(FF) 31 -(FF) 32 -(FF)<舍 33 -(FF) 34 -(FF) 35 -(FF) 36 -(FF) 37 -(FF) 38 -(FF) 39 -(FF) 40 -(FF) Note: There seems to be a mistake in the original text where "L" appears in Formula XXXII (ID=40). It should probably be a number or a correct symbol. Also, "舍0001664" in ID=87 seems incorrect. I've translated it as it is but it should be verified.41 -(FF) 42 -(FF) 43 -(FF) 44 -(FF) 45 (Formula XXXII) And (FF)1-(FF) 45 Their identities are provided in Table 33; Where equation XXXIII is represented as: (GG)1-(GG)2-(GG)3-(GG)4-(GG)5-(GG)6-(GG)7-(GG)8-(GG)9-(GG) 10 -(GG) 11 -(GG) 12 -(GG) 13 -(GG) 14 -(GG) 15 -(GG) 16 -(GG) 17 -(GG) 18 -(GG) 19 -(GG) 20 -(GG) 21 -(GG) 22 -(GG) 23 -(GG) 24 -(GG) 25 -(GG) 26 -(GG) 27 -(GG) 28 -(GG) 29 -(GG) 30 -(GG) 31 -(GG) 32 -(GG) 33 -(GG) 34 -(GG) 35 -(GG) 36 -(GG) 37 -(GG) 38 -(GG) 39 -(GG) 40 -(GG) 41 -(GG) 42 -(GG) 43 -(GG) 44 -(GG) 45 -(GG) 46 (Formula XXXIII) And (GG)1-(GG) 46 Their identities are provided in Table 34; Wherein, XXXIV is represented as: (HH)1-(HH)2-(HH)3-(HH)4-(HH)5-(HH)6-(HH)7-(HH)8-(HH)9-(HH) 10 -(HH) 11 -(HH) 12 -(HH) 13 -(HH) 14 -(HH) 15 -(HH) 16 -(HH) 17 -(HH) 18 -(HH) 19 -(HH) 20 -(HH) 21 -(HH) 22 -(HH) 23 -(HH) 24 -(HH) 25 -(HH) 26 -(HH) 27 -(HH) 28 -(HH) 29 -(HH) 30 -(HH) 31 -(HH) 32 -(HH) 33 -(HH) 34 -(HH) 35 -(HH) 36 -(HH) 37 -(HH) 38 -(HH) 39 -(HH) 40 -(HH) 41 -(HH) 42 -(HH) 43 -(HH) 44 -(HH) 45 -(HH) 46 -(HH) 47 (Formula XXXIV) and (HH)1-(HH) 47 has the identity as provided in Table 35; where Formula XXXV is represented as: (II)1-(II)2-(II)3-(II)4-(II)5-(II)6-(II)7-(II)8-(II)9-(II) 10 -(II) 11 -(II) 12 -(II) 13 -(II) 14 -(II) 15 -(II)16 -(II) 17 -(II) 18 -(II) 19 -(II) 20 -(II) 21 -(II) 22 -(II) 23 -(II) 24 -(II) 25 -(II) 26 -(II) 27 -(II) 28 -(II) 29 -(II) 30 -(II) 31 -(II) 32 -(II) 33 -(II) 34 -(II) 35 -(II) 36 -(II) 37 -(II) 38 -(II) 39 -(II) 40 -(II) 41 -(II) 42 -(II) 43 -(II) 44 -(II) 45 -(II) 46 -(II) 47 -(II) 48 (Formula XXXV) And (II)1-(II) 48 The identities are provided in Table 36; Wherein, XXXVI is represented as: (JJ)1-(JJ)2-(JJ)3-(JJ)4-(JJ)5-(JJ)6-(JJ)7-(JJ)8-(JJ)9-(JJ) 10 -(JJ) 11 -(JJ) 12 -(JJ) 13 -(JJ) 14 -(JJ) 15 -(JJ) 16 -(JJ) 17 -(JJ) 18 -(JJ) 19 -(JJ) 20 -(JJ) 21 -(JJ) 22 -(JJ) 23 -(JJ)24 -(JJ) 25 -(JJ) 26 -(JJ) 27 -(JJ) 28 -(JJ) 29 -(JJ) 30 -(JJ) 31 -(JJ) 32 -(JJ) 33 -(JJ) 34 -(JJ) 35 -(JJ) 36 -(JJ) 37 -(JJ) 38 -(JJ) 39 -(JJ) 40 -(JJ) 41 -(JJ) 42 -(JJ) 43 -(JJ) 44 -(JJ) 45 -(JJ) 46 -(JJ) 47 -(JJ) 48 -(JJ) 49 (Formula XXXVI) And (JJ)1-(JJ) 49 Their identities are provided in Table 37; Where equation XXXVII is represented as: (KK)1-(KK)2-(KK)3-(KK)4-(KK)5-(KK)6-(KK)7-(KK)8-(KK)9-(KK) 10 -(KK) 11 -(KK) 12 -(KK) 13 -(KK) 14 -(KK) 15 -(KK) 16 -(KK) 17 -(KK) 18 -(KK) 19 -(KK) 20 -(KK) 21 -(KK) 22 -(KK) 23 -(KK) 24 -(KK) 25 -(KK) 26 -(KK) 27 -(KK) 28 -(KK) 29 -(KK) 30 -(KK)31 -(KK) 32 -(KK) 33 -(KK) 34 -(KK) 35 -(KK) 36 -(KK) 37 -(KK) 38 -(KK) 39 -(KK) 40 -(KK) 41 -(KK) 42 -(KK) 43 -(KK) 44 -(KK) 45 -(KK) 46 -(KK) 47 -(KK) 48 -(KK) 49 -(KK) 50 -(KK) 51 -(KK) 52 (Formula XXXVII) And (KK)1-(KK) 52 The identities are provided in Table 38; Wherein, equation XXXVIII is represented as: (LL)1-(LL)2-(LL)3-(LL)4-(LL)5-(LL)6-(LL)7-(LL)8-(LL)9-(LL) 10 -(LL) 11 -(LL) 12 -(LL) 13 -(LL) 14 -(LL) 15 -(LL) 16 -(LL) 17 -(LL) 18 -(LL) 19 -(LL) 20 -(LL) 21 -(LL) 22 -(LL) 23 -(LL) 24 -(LL) 25 -(LL) 26 -(LL) 27 -(LL) 28 -(LL) 29 -(LL) 30 -(LL) 31 -(LL) 32 -(LL) 33 -(LL) 34 -(LL)35 -(LL) 36 -(LL) 37 -(LL) 38 -(LL) 39 -(LL) 40 -(LL) 41 -(LL) 42 -(LL) 43 -(LL) 44 -(LL) 45 -(LL) 46 -(LL) 47 -(LL) 48 -(LL) 49 -(LL) 50 -(LL) 51 -(LL) 52 -(LL) 53 -(LL) 54 -(LL) 55 (Formula XXXVIII) and (LL)1 - (LL) 55 the identity of which is provided in Table 39; and where Formula XXXIX is represented as: (MM)1 - (MM)2 - (MM)3 - (MM)4 - (MM)5 - (MM)6 - (MM)7 - (MM)8 - (MM)9 - (MM) 10 -(MM) 11 -(MM) 12 -(MM) 13 -(MM) 14 -(MM) 15 -(MM) 16 -(MM) 17 -(MM) 18 -(MM) 19 -(MM) 20 -(MM) 21 -(MM) 22 -(MM) [[ID=七十六]] 23 [[ID=七十七]]-(MM) [[ID=七十八]] 24 [[ID=七十九]]-(MM) [[ID=八十]] 25 [[ID=八十一]]-(MM) [[ID=八十二]] 26 [[ID=八十三]]-(MM) [[ID=八十四]] 27 [[ID=八十五]]-(MM) [[ID=八十六]] 28 [[ID=八十七]]-(MM) [[ID=八十八]] 29 [[ID=八十九]]-(MM) [[ID=九十]] 30 [[ID=九十一]]-(MM) [[ID=九十二]] 31 [[ID=九十三]]-(MM) [[ID=九十四]] 32 [[ID=九十五]]-(MM) [[ID=九十六]] 33 [[ID=九十七]]-(MM) [[ID=九十八]] 34 [[ID=九十九]]-(MM) [[ID=一百]] 35 [[ID=一百零一]]-(MM)36 -(MM) 37 -(MM) 38 -(MM) 39 -(MM) 40 -(MM) 41 -(MM) 42 -(MM) 43 -(MM) 44 -(MM) 45 -(MM) 46 -(MM) 47 -(MM) 48 -(MM) 49 -(MM) 50 -(MM) 51 -(MM) 52 -(MM) 53 -(MM) 54 -(MM) 55 -(MM) 56 -(MM) 57 -(MM) 58 -(MM) 59 (Formula XXXIX) And (MM)1-(MM) 59 Their identities are provided in Table 40.
[0012] In some embodiments, the signal peptide X1 comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 99, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
[0013] In some implementations, the payload protein is a therapeutic peptide or protein.
[0014] In some embodiments, a nucleic acid molecule is provided. In some embodiments, the nucleic acid molecule encodes a signal peptide as provided herein.
[0015] In some embodiments, a nucleic acid molecule is provided. In some embodiments, the nucleic acid molecule encodes a recombinant polypeptide as provided herein.
[0016] In some embodiments, a vector is provided. In some embodiments, the vector comprises a nucleic acid molecule as provided herein. In some embodiments, the nucleic acid molecule encodes a signal peptide as provided herein. In some embodiments, the nucleic acid molecule encodes a recombinant polypeptide as provided herein.
[0017] In some embodiments, a cell is provided. In some embodiments, the cell comprises nucleic acid molecules as provided herein. In some embodiments, the cell comprises a vector as provided herein. In some embodiments, the nucleic acid molecule encodes a signal peptide as provided herein. In some embodiments, the nucleic acid molecule encodes a recombinant polypeptide as provided herein.
[0018] In some embodiments, a composition is provided. In some embodiments, the composition comprises a nucleic acid molecule as provided herein. In some embodiments, the composition comprises a vector as provided herein. In some embodiments, the nucleic acid molecule encodes a signal peptide as provided herein. In some embodiments, the nucleic acid molecule encodes a recombinant polypeptide as provided herein.
[0019] In some embodiments, a method is provided for treating a disease or condition in a subject of need. In some embodiments, the method includes administering to the subject an effective amount of a nucleic acid molecule, as provided herein, thereby treating the disease or condition. In some embodiments, the nucleic acid molecule encodes a recombinant polypeptide, as provided herein. In some embodiments, the disease or condition is cancer.
[0020] In some embodiments, a method for treating cancer in a subject of need is provided. In some embodiments, the method includes administering to the subject a carrier comprising a nucleic acid molecule encoding a signal peptide fused to or linked with a payload protein, wherein the signal peptide is a signal peptide as provided herein, and wherein the payload protein is a therapeutic peptide or protein that can be used to treat cancer. Attached Figure Description
[0021] Figure 1A A schematic diagram of an mCherry containing an N-terminus signal peptide is provided, and its use for signal peptide screening via in vitro pDNA transfection is shown.
[0022] Figure 1B Representative images of mCherry tagged with a signal peptide co-localized with CellMask green plasma membrane staining agent are shown. Top image: SP is Nov-Mem-18-1 (SEQ ID NO: 5). Bottom image: SP is Nov-Mem-21-1 (SEQ ID NO: 8).
[0023] Figure 1CRepresentative images of mCherry tagged with a signal peptide co-localized with CellMask green plasma membrane staining agent are shown. Top image: SP is Nov-Mem-19-1 (SEQ ID NO: 6). Bottom image: SP is Nov-Mem-22-1 (SEQ ID NO: 9).
[0024] Figure 1D Representative images of mCherry tagged with a signal peptide co-localized with CellMask green plasma membrane staining agent are shown. Top panel: SP is Nov-Mem-20-1 (SEQ ID NO: 7). Bottom panel: No SP control.
[0025] Figure 1E Examples are given corresponding to Figure 1B , Figure 1C and Figure 1D Quantitative analysis of representative images from experiments.
[0026] Figure 1F Examples are given corresponding to Figure 1B , Figure 1C and Figure 1D Quantitative analysis of representative images from experiments.
[0027] Figure 2A Quantification of mCherry fluorescence in HeLa cell lysates and culture medium at different time points was provided. mCherry without the signal peptide was used as a control (“WT-mCherry”). gLuc, Gaussian luciferase.
[0028] Figure 2B The results provided quantification of mCherry fluorescence in HeLa cell lysates and culture medium after 72 hours.
[0029] Figure 2C Quantification of mCherry fluorescence in Huh7 cell lysates and culture medium was provided 72 h post-transfection.
[0030] Figure 3A We provided quantification of time-dependent (100 ng mRNA / well) and dose-dependent (at 72 h) mCherry secretion in Huh7 cells, culture medium, and cell lysates.
[0031] Figure 3B The quantification of mCherry signaling in different cell lines is shown. Cells in 96-well plates were treated with mDLNP-mRNA at given time points. Detailed Implementation
[0032] Before describing the compositions and methods of the present invention, it should be understood that the scope of this disclosure is not limited to the specific processes, compositions, or methodologies described herein, as these are subject to change. It should also be understood that the terminology used herein is for the purpose of describing a particular version or embodiment only and is not intended to limit the scope of this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. It should be further understood that any methods and materials similar to or equivalent to those described and materials herein can be used in the practice or testing of embodiments of the methods and systems disclosed herein, and such equivalents are within the scope of this disclosure.
[0033] definition The following explanations of terms and methods are provided to better describe this disclosure and to guide those skilled in the art in practicing this disclosure.
[0034] As used herein, unless the context explicitly indicates otherwise, "comprising" means "including" and the singular forms "a," "an," or "the" include a plural number of indicators. For example, references to "comprising a therapeutic agent" include one or more such therapeutic agents. Unless the context explicitly indicates otherwise, the term "or" refers to a single element among the stated alternative elements. For example, the phrase "A or B" means A alone or B alone. The phrase "A, B, or a combination thereof" means A alone, B alone, or a combination of A and B. Similarly, "one or more of A and B" means A, B, or a combination of both A and B. The phrase "A and B" means a combination of A and B. Furthermore, the various elements, features, and steps discussed herein, as well as other known equivalents of each such element, feature, or step, can be mixed and matched by those skilled in the art to perform methods according to the principles described herein. Among the various elements, features, and steps, some will be specifically included in a particular instance, while others will be specifically excluded from a particular instance.
[0035] In some instances, figures used to describe and claim certain embodiments, representing the amount of an ingredient, properties such as molecular weight, reaction conditions, etc., should be understood to be modified in some cases by the terms “about” or “approximately.” For example, “about” or “approximately” can indicate a variation of + / - 10%, + / - 5%, or + / - 1% in the value it describes. Thus, in some embodiments, the numerical parameters set forth herein are approximate values that can vary depending on the desired characteristics of a particular embodiment. Furthermore, unless the context indicates otherwise, when the phrase enumerates “about x to y,” the term “about” modifies both x and y and can be used interchangeably with the phrase “about x to about y.”
[0036] As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing exogenous nucleic acids into host cells, including calcium phosphate or calcium chloride coprecipitation, DEAE-dextran-mediated transfection, lipid transfection (e.g., using commercially available reagents such as, for example, LIPOFECTIN® (Invitrogen Corp., San Diego, CA), LIPOFECTAMINE® (Invitrogen), FUGENE® (Roche Applied Science, Basel, Switzerland), JETPEI™ (Polyplus-transfection Inc., New York, NY), EFFECTENE® (Qiagen, Valencia, CA), DREAMFECT™ (OZ Biosciences, France)), or electroporation (e.g., in vivo electroporation). Suitable methods for transforming or transfecting host cells can be found in Sambrook et al. ( Molecular Cloning: A Laboratory Manual. (2nd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989) and other laboratory manuals.
[0037] Methods and materials for non-viral delivery of nucleic acids to cells also include biolistics, virions, liposomes, lipid nanoparticles, immunoliposomes, polycationic or lipid-nucleic acid conjugates, naked DNA, artificial viral particles, and drug-enhanced DNA uptake. Lipid transfection is described in U.S. Patent Nos. 5,049,386, 4,946,787, and 4,897,355, and lipid transfection reagents are commercially available (e.g., TRANSFECTAM™ and LIPOFECTIN™). Cationic and neutral lipids suitable for effective receptor recognition of polynucleotides in lipid transfection include those disclosed in WO 91 / 17424 and WO 91 / 16024.
[0038] The chemical composition of peptides will be described herein by a series of single-letter abbreviations of amino acids, or “amino acid sequence” or “sequence”, which are conventional and known to those skilled in the art. Although a reference sequence will be explicitly disclosed, it may be modified in any aspect and embodiment to include conserved amino acid substitutions, as well as variants and fragments, while maintaining the characteristics and functionality of the reference sequence.
[0039] As used herein, a "payload protein" or "protein of interest" refers to a protein that will be generated by the host cell and chaperoned to a desired intracellular location (e.g., chaperoned to the cell membrane). After translocation to the appropriate intracellular space, all, some, or no novel signal peptides may be fused to the payload protein. Optionally, the payload protein, still partially or completely linked to the novel signal peptide, may be further processed, for example, to remove any remaining novel signal peptides. The payload protein can be any protein known or yet to be known, such as enzymes, enzyme inhibitors, growth factors, hormones, antibodies, antigens, vaccines, therapeutics, or any combination thereof. More specific examples are given below.
[0040] As used herein, the terms “fused” or “linked” when referring to proteins with different domains or heterologous sequences mean that the protein domains are parts of the same peptide chain linked together by peptide bonds or other covalent bonds. Domains or segments may be directly linked or fused to each other, or another domain or peptide sequence may be between two domains or sequences, and such sequences will still be considered fused or linked together. In some embodiments, the various domains or proteins provided herein are directly linked or fused to each other or with adapter sequences, such as the glycine / serine sequence described herein that links two domains together.
[0041] As used herein, “identity” refers to the subunit sequence identity between two polymeric molecules, such as two nucleic acid or amino acid molecules, or between two polynucleotide or polypeptide molecules. Two amino acid sequences are identical when they have the same residue at the same position; for example, if each of two polypeptide molecules has an arginine residue at that position, then they are identical at that position. The degree of identity, or similarity, between two amino acid or two nucleic acid sequences having the same residue at the same position in an alignment is usually expressed as a percentage. Identity between two amino acid or two nucleic acid sequences is a direct function of the number of matching or identical positions; for example, if half of the positions in two sequences are identical, then the two sequences have 50% identity; if 90% of the positions (e.g., 9 out of 10) match or are identical, then the two amino acid sequences have 90% identity.
[0042] "Substantially identical" means that the polypeptide or nucleic acid molecule has at least 50% identity with a reference amino acid sequence (e.g., any of the amino acid sequences described herein) or nucleic acid sequence (e.g., any of the nucleic acid sequences described herein). In some embodiments, this sequence has at least 60%, 80%, or 85%, or 90%, 95%, or even 99% identity with the sequence used for comparison at the amino acid level or nucleic acid level. Other percentages of identity with respect to a particular sequence are described herein.
[0043] Sequence identity can be measured / determined using sequence analysis software (e.g., sequence analysis packages from the Genetics Computing Group of the University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, including BLAST, BESTFIT, GAP, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTALOMEGA, MUSCLE, or PILEUP / PRETTYBOX programs). Such software matches identical or similar sequences by specifying degrees of homology for various substitutions, deletions, and / or other modifications. Conserved substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In exemplary methods for determining the degree of identity, a BLAST program can be used, where a probability score between e3 and e100 indicates closely related sequences. In some embodiments, sequence identity is determined using BLAST with default settings. In some implementations, Clustal Omega is used to determine sequence identity.
[0044] For the purposes of the embodiments provided herein, compositions comprising various proteins may be included, in some cases, containing amino acid sequences that have sequence identity with the amino acid sequences disclosed herein. Therefore, in some embodiments, depending on the specific sequence, the degree of sequence identity with the SEQ ID NO disclosed herein is preferably greater than 50% (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher). In addition to these percentages, other percentages of identity are also provided herein. Identity between peptides can be determined using an affine gap search with parameters vacancy opening penalty – 12 and vacancy extension penalty = 1, as implemented in the MPSRCH (Oxford Molecular) program.
[0045] Compared to publicly disclosed proteins, these proteins may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) conserved amino acid substitutions, i.e., one amino acid is replaced by another amino acid with the relevant side chain. Genetically encoded amino acids are generally classified into four families: (1) acidic, i.e., aspartic acid and glutamic acid; (2) basic, i.e., lysine, arginine, and histidine; (3) nonpolar, i.e., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and (4) uncharged polar, i.e., glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Phenylalanine, tryptophan, and tyrosine are sometimes collectively classified as aromatic amino acids. Generally, the substitution of a single amino acid within these families does not have a significant impact on biological activity. Proteins may have one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) single amino acid deletions relative to publicly disclosed protein sequences. Proteins may also include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) insertions relative to a published protein sequence (e.g., each of 1, 2, 3, 4, or 5 amino acids).
[0046] "Encoding" refers to the inherent property of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA, or mRNA) to serve as a template for the synthesis of other polymers and macromolecules in biological processes. These polymers and macromolecules have defined nucleotide sequences (i.e., rRNA, tRNA, and mRNA) or defined amino acid sequences, and the resulting biological properties. Therefore, if the transcription and translation of the mRNA corresponding to a gene produces a protein in a cell or other biological system, then that gene encodes a protein. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in sequence listings, and the non-coding strand, which serves as a template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.
[0047] As used in this article, the following abbreviations for common nucleic acid bases are used: "A" refers to adenosine, "C" refers to cytosine, "G" refers to guanosine, "T" refers to thymidine, and "U" refers to uridine.
[0048] Unless otherwise specified, "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate versions of each other and encode the same amino acid sequence. A phrase encoding a protein or RNA nucleotide sequence may also include introns, provided that the nucleotide sequence encoding that protein may contain introns in some versions.
[0049] The term "oligonucleotide" generally refers to short polynucleotides. It should be understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, C, G), this also provides a corresponding RNA sequence (i.e., A, U, C, G), where "U" replaces "T".
[0050] As used herein, the term "polynucleotide" is defined as a nucleotide chain. Furthermore, nucleic acids are polymers of nucleotides. Therefore, as used herein, the terms "nucleic acid" or "nucleic acid molecule" and "polynucleotide" are interchangeable. As used herein, polynucleotides include, but are not limited to, all nucleic acid sequences obtained by any method available in the art and by synthetic means, including but not limited to recombinant methods, i.e., cloning nucleic acid sequences from recombinant libraries or cell genomes using cloning techniques such as PCR.
[0051] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably and refer to compounds comprising a plurality of amino acid residues covalently linked by peptide bonds. As used herein, the term refers to short chains, which are also commonly referred to in the art as, for example, peptides, oligopeptides, and oligomers; and long chains, which are commonly referred to in the art as proteins, which exist in many types. “Polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, etc. Polypeptides include natural peptides, recombinant peptides, synthetic peptides, or combinations thereof.
[0052] As used herein, unless otherwise indicated, "antibody fragment" or "antigen-binding fragment" means an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to the antigen to which the full-length antibody is bound, such as a fragment retaining one or more CDR regions. Examples of antibody-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; bivalent antibodies; linear antibodies; single-chain antibody molecules, such as sc-Fv; nanobodies (single-domain antibodies); and multispecific antibodies formed from antibody fragments.
[0053] The "Fab fragment" contains a light chain and a heavy chain of C. H 1. Variable region. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule.
[0054] The “Fc” region contains two C-cells containing antibodies. H 2 and C H 3. Heavy chain segments of structural domains. Two heavy chain segments are connected by two or more disulfide bonds and by C... H The hydrophobic interactions of the three structural domains are combined.
[0055] A “Fab” fragment contains a portion or segment of a light chain and a heavy chain, the portion or segment containing VH Domain and C H 1. Structural domains and C H 1 and C H The region between the two structural domains allows interchain disulfide bonds to form between the two heavy chains of the two Fab' segments to form the F(ab')2 molecule.
[0056] The “F(ab')2 fragment” contains two light chains and two chains containing C. H 1 and C H The heavy chains, which constitute a portion of the constant region between the two heavy chains, allow interchain disulfide bonds to form between them. Therefore, the F(ab')2 segment consists of two Fab' segments bonded together by disulfide bonds between the two heavy chains.
[0057] The “Fv region” contains variable regions from both the heavy and light chains, but lacks constant regions.
[0058] The term "single-chain Fv" or "scFv" antibody refers to a V antibody containing an antibody. H and V L Antibody fragments containing domains, wherein these domains are present within a single polypeptide chain. Generally, Fv polypeptides also contain V... H With V L A peptide linker between the domains allows scFv to form the structure required for antigen binding. For a review of scFv, see Pluckthun (1994). Volume 113, edited by Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315. See also International Patent Application Publication No. WO 88 / 01649 and U.S. Patent Nos. 4,946,778 and 5,260,203.
[0059] Antibody molecules can be monospecific (e.g., monovalent or bivalent), bispecific (e.g., bivalent, trivalent, tetravalent, pentavalent, or hexavalent), trispecific (e.g., trivalent, tetravalent, pentavalent, or hexavalent), or have a higher order of specificity (e.g., tetraspecific) and / or a higher order of valence than hexavalent. Antibody molecules can contain functional fragments of both the light chain variable region and the heavy chain variable region, or the heavy and light chains can be fused together to form a single polypeptide.
[0060] As used herein, the terms “comprising” (and any form of inclusion, such as “comprise”, “comprises”, and “comprised”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unlisted elements or method steps. Any step or composition using the transitional phrase “comprise” or “comprising” can also be described using the transitional phrase “consisting of” or “consists”.
[0061] As used herein, the term “contact” means bringing together two elements of an in vitro or in vivo system. For example, “contacting” a virus or vector described herein with an individual or patient or cell includes administering the virus to an individual or patient (such as a human), and, for example, introducing a compound into a sample containing a cell preparation or a purified preparation containing such cells.
[0062] As used herein, the interchangeable terms “individual” or “subject” or “patient” mean any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates such as humans. In some embodiments, the subject is a human. A “subject in need” means a subject who has been identified as requiring treatment for a condition to be treated and is being treated with the specific intent to treat such condition. A condition can be any condition described herein, for example.
[0063] The compositions disclosed herein can be administered to a subject in a variety of ways by administering the composition to the subject. As used herein, administering or administering means providing or providing the composition to a subject. As used herein, oral administration means delivery of the active agent through the mouth. As used herein, topical administration means delivery of the active agent to the body surface, such as the skin, mucous membranes (e.g., nasal membranes, vaginal membranes, buccal membranes, etc.).
[0064] "Disease" is a health condition in which an animal is unable to maintain homeostasis, and if the disease is not treated, the animal's health continues to deteriorate. In contrast, an animal's "symptom" is a health condition in which the animal is able to maintain homeostasis, but the animal's health is not as good as it would be in the absence of the symptom. A symptom, if left untreated, does not necessarily lead to further deterioration of the animal's health.
[0065] The terms "effective amount" or "therapeutic effective amount" are used interchangeably herein and refer to the amount of a compound, formulation, material, or composition as described herein that effectively achieves a particular biological outcome or provides a therapeutic or preventative benefit. Such outcomes may include, but are not limited to, the amount of immune cell activation induced at a detectable level when administered to a mammal, compared to immune cell activation detected in the absence of the composition. Immune responses can be readily assessed using a wide range of methods recognized in the art. Those skilled in the art will understand that variations in the amount of the composition administered herein can be readily determined based on a number of factors, such as the disease or symptom being treated, the age and health status and physical condition of the mammal being treated, the severity of the disease, and the specific compound administered.
[0066] Scope: Throughout this disclosure, various aspects of the embodiments may be presented in the form of scope. It should be understood that the description in scope form is merely for convenience and brevity and should not be construed as an immutable limitation. Therefore, the description of scope should be considered as having specifically disclosed all possible sub-scopes and individual numerical values within that scope. For example, a description of a scope such as 1 to 6 should be considered as having specifically disclosed sub-scopes such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numbers within that scope, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the width of the scope. Unless otherwise explicitly stated to the contrary, the disclosed scope also includes the endpoints of the scope.
[0067] Novel signal peptides Unbound by any particular theory, the embodiments provided herein have been found to demonstrate that novel signal peptides can guide the expression of payload proteins, as described herein, at the cell membrane. The payload proteins may or may not naturally contain a signal peptide that guides the protein to a specific cellular compartment (e.g., the cell membrane). Therefore, the novel signal peptides provided can guide the enhanced expression of native membrane proteins to the cell membrane, or can guide payload proteins (e.g., therapeutic proteins) to the cell membrane for expression.
[0068] In some embodiments, a signal peptide is provided. In some embodiments, the signal peptide comprises the amino acid sequences listed in Table 1 below: Table 1: Exemplary Novel Signal Peptides
[0069] In some embodiments, the signal peptide comprises an amino acid sequence substantially similar to the amino acid sequences listed in Table 1. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequences listed in Table 1. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 81, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39) of the sequence. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 2. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 3. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 4. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 5.In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 6. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 7. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 8. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 9. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 10. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 11. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 12. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 13.In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 14. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 15. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 16. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 17. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 18. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 19. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 20. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 21.In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 22. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 23. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 24. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 25. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 26. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 27. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 28. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 29.In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 30. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 31. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 32. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 33. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 34. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 35. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 36. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 37.In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 38. In some embodiments, the signal peptide comprises an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identity with the amino acid sequence of SEQ ID NO: 39.
[0070] In some embodiments, the signal peptide is a variant of the amino acid sequence provided in Table 1. In some embodiments, the signal peptide comprises an amino acid sequence as defined by formulas I-XXXIX below.
[0071] Variants of SEQ ID NO: 1 (Formula I) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: A1-A2-A3-A4-A5-A6-A7-A8-A9-A 10 -A 11 -A 12 -A 13 -A 14 (Formula I); Where A1–A 14 The identities of each of them are provided in Table 2 below.
[0072] Table 2
[0073] In some embodiments, A1 is methionine (M). In some embodiments, A2 is an amino acid selected from the group consisting of L, G, P, V, and W. In some embodiments, A3 is an amino acid selected from the group consisting of L, A, W, and A. In some embodiments, A4 is an amino acid selected from the group consisting of L, R, E, and S. In some embodiments, A5 is an amino acid selected from the group consisting of L, F, S, and A. In some embodiments, A6 is an amino acid selected from the group consisting of L, F, and P. In some embodiments, A7 is an amino acid selected from the group consisting of L, F, and C. In some embodiments, A8 is an amino acid selected from the group consisting of L, A, and V. In some embodiments, A9 is an amino acid selected from the group consisting of L, V, and P. In some embodiments, A... 10 These are amino acids selected from the group consisting of L, T, I, A, and G. In some embodiments, A... 11These are amino acids selected from the group consisting of S, F, A, V, G, and L. In some implementations, A... 12 These are amino acids selected from the group consisting of A, G, L, I, and D. In some embodiments, A... 13 These are amino acids selected from the group consisting of A, R, H, F, T, and G. In some implementations, A... 14 It is an amino acid group selected from A, S, P and Q.
[0074] It should be understood that variants of SEQ ID NO: 1 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula I. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 2 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 3 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 4 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 5 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 6 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 7 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and the substitution at position 8 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 9 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 10 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 11 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 12 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 13 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at position 14 of SEQ ID NO: 1. Variations may also comprise any number of substitutions provided by Formula I. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 of SEQ ID NO: 1. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 1 and a substitution at more than one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 of SEQ ID NO: 1.
[0075] Variants of SEQ ID NO: 2 (Formula II) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: B1-B2-B3-B4-B5-B6-B7-B8-B9-B 10 -B 11 -B 12 -B 13 -B 14 -B 15 (Formula II); Where B1–B 15 The identities of each of them are provided in Table 3 below.
[0076] Table 3
[0077] In some embodiments, B1 is methionine (M). In some embodiments, B2 is an amino acid selected from the group consisting of L, A, F, and R. In some embodiments, B3 is an amino acid selected from the group consisting of L, V, A, and F. In some embodiments, B4 is leucine (L). In some embodiments, B5 is an amino acid selected from the group consisting of L, V, and A. In some embodiments, B6 is an amino acid selected from the group consisting of L, F, I, and T. In some embodiments, B7 is an amino acid selected from the group consisting of L, S, and I. In some embodiments, B8 is an amino acid selected from the group consisting of L, C, and V. In some embodiments, B9 is an amino acid selected from the group consisting of L and W. In some embodiments, B... 10 These are amino acids selected from the group consisting of L, V, A, and S. In some implementations, B... 11 These are amino acids selected from the group consisting of G, L, P, V, and W. In some implementations, B... 12 These are amino acids selected from the group consisting of S, L, M, V, W, and P. In some implementations, B... 13 These are amino acids selected from the group consisting of V, L, I, G, and A. In some implementations, B... 14 These are amino acids selected from the group consisting of L, T, A, and E. In some implementations, B... 15 These are amino acids selected from the group consisting of G, S, and A.
[0078] It should be understood that variants of SEQ ID NO: 2 do not need to include a substitution at every position providing the substitution, but only one substitution within the scope of Formula II. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 2 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 3 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 5 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 6 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 7 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 8 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and the substitution at position 9 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 10 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 11 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 12 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 13 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 14 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at position 15 of SEQ ID NO: 2. Variations may also comprise any number of substitutions provided by Formula II. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at at least one position corresponding to position 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of SEQ ID NO: 2. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 2 and a substitution at more than one position corresponding to position 2, 3, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of SEQ ID NO: 2.
[0079] Variants of SEQ ID NO: 3 (Formula III) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: C1-C2-C3-C4-C5-C6-C7-C8-C9-C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 (Formula III); Where C1–C 16 The identities of each of them are provided in Table 4 below.
[0080] Table 4
[0081] In some embodiments, C1 is methionine (M). In some embodiments, C2 is an amino acid selected from the group consisting of L, A, and V. In some embodiments, C3 is an amino acid selected from the group consisting of L and P. In some embodiments, C4 is an amino acid selected from the group consisting of L and A. In some embodiments, C5 is an amino acid selected from the group consisting of L and V. In some embodiments, C6 is leucine (L). In some embodiments, C7 is leucine (L). In some embodiments, C8 is leucine (L). In some embodiments, C9 is leucine (L). In some embodiments, C... 10 It is leucine (L). In some implementations, C 11 These are amino acids selected from the group consisting of L and V. In some embodiments, C 12 These are amino acids selected from the group consisting of G, L, P, and S. In some implementations, C... 13 These are amino acids selected from the group consisting of L and G. In some embodiments, C... 14 These are amino acids selected from the group consisting of A, C, L, S, and V. In some embodiments, C... 15 It is leucine (L). In some implementations, C 16 These are amino acids selected from the group consisting of A, G, and S.
[0082] It should be understood that variants of SEQ ID NO: 3 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula III. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 2 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 3 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 4 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 5 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 11 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 12 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and the substitution at position 13 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and a substitution at position 14 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and a substitution at position 16 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and a substitution at position 17 of SEQ ID NO: 3. Variants may also comprise any number of substitutions provided by Formula III. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 11, 12, 13, 14, or 16 of SEQ ID NO: 3. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 3 and a substitution at more than one position corresponding to positions 2, 3, 4, 5, 11, 12, 13, 14, or 16 of SEQ ID NO: 3.
[0083] Variants of SEQ ID NO: 4 (Formula IV) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: D1-D2-D3-D4-D5-D6-D7-D8-D9-D 10 -D 11 -D 12 -D 13 -D14 -D 15 -D 16 -D 17 (Form IV); Where D1–D 17 The identities of each of them are provided in Table 5 below.
[0084] Table 5
[0085] In some embodiments, D1 is methionine (M). In some embodiments, D2 is an amino acid selected from the group consisting of L, A, and W. In some embodiments, D3 is leucine (L). In some embodiments, D4 is leucine (L). In some embodiments, D5 is leucine (L). In some embodiments, D6 is an amino acid selected from the group consisting of L, A, and W. In some embodiments, D7 is leucine (L). In some embodiments, D8 is leucine (L). In some embodiments, D9 is leucine (L). In some embodiments, D... 10 These are amino acids selected from the group consisting of L and A. In some implementations, D... 11 These are amino acids selected from the group consisting of L, F, and S. In some implementations, D... 12 These are amino acids selected from the group consisting of L, A, and F. In some implementations, D... 13 These are amino acids selected from the group consisting of G, P, and S. In some implementations, D... 14 These are amino acids selected from the group consisting of L, P, S, and A. In some implementations, D... 15 These are amino acids selected from the group consisting of A, V, and T. In some implementations, D... 16 These are amino acids selected from the group consisting of L, S, and A. In some implementations, D... 17 These are amino acids selected from the group consisting of A, G, and S.
[0086] It should be understood that variants of SEQ ID NO: 4 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula IV. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 2 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 6 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 10 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 11 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 12 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 13 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and the substitution at position 14 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and a substitution at position 15 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and a substitution at position 16 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and a substitution at position 17 of SEQ ID NO: 4. Variations may also comprise any number of substitutions provided by Formula IV. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and a substitution at at least one position corresponding to positions 2, 6, 10, 11, 12, 13, 14, 15, 16, or 17 of SEQ ID NO: 4. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 4 and substitutions at more than one position corresponding to positions 2, 6, 10, 11, 12, 13, 14, 15, 16, or 17 of SEQ ID NO: 4.
[0087] Variant of SEQ ID NO: 5 (Formula V) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: E1-E2-E3-E4-E5-E6-E7-E8-E9-E 10 -E 11 -E 12 -E13 -E 14 -E 15 -E 16 -E 17 -E 18 (Formula V); Where E1–E 18 The identities of each of them are provided in Table 6 below.
[0088] Table 6
[0089] In some embodiments, E1 is methionine (M). In some embodiments, E2 is an amino acid selected from the group consisting of A, R, and L. In some embodiments, E3 is an amino acid selected from the group consisting of L, P, and M. In some embodiments, E4 is an amino acid selected from the group consisting of L, V, and A. In some embodiments, E5 is leucine (L). In some embodiments, E6 is an amino acid selected from the group consisting of L, V, and A. In some embodiments, E7 is an amino acid selected from the group consisting of L and F. In some embodiments, E8 is leucine (L). In some embodiments, E9 is leucine (L). In some embodiments, E... 10 These are amino acids selected from the group consisting of L and I. In some embodiments, E... 11 These are amino acids selected from the group consisting of L and A. In some implementations, E... 12 It is leucine (L). In some implementations, E 13 These are amino acids selected from the group consisting of L, I, F, and V. In some implementations, E... 14 These are amino acids selected from the group consisting of L, P, and G. In some implementations, E... 15 These are amino acids selected from the group consisting of G and L. In some implementations, E... 16 These are amino acids selected from the group consisting of V, A, L, S, and T. In some embodiments, E... 17 These are amino acids selected from the group consisting of S, L, and A. In some implementations, E... 18 These are amino acids selected from the group consisting of A, G, and S.
[0090] It should be understood that variants of SEQ ID NO: 5 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula V. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 2 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 3 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 4 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 6 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 7 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 10 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and the substitution at position 11 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 13 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 14 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 15 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 16 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 17 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at position 18 of SEQ ID NO: 5. Variations may also comprise any number of substitutions provided by Formula V. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at at least one position corresponding to position 2, 3, 4, 6, 7, 10, 11, 13, 14, 15, 16, 17, or 18 of SEQ ID NO: 5. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 5 and a substitution at more than one position corresponding to position 2, 3, 4, 6, 7, 10, 11, 13, 14, 15, 16, 17, or 18 of SEQ ID NO: 5.
[0091] Variants of SEQ ID NO: 6 (Formula VI) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: F1-F2-F3-F4-F5-F6-F7-F8-F9-F 10 -F 11 -F 12 -F 13 -F 14 -F 15 -F 16 -F 17 -F 18 -F 19 (Formula VI); Where F1–F 19 The identities of each of them are provided in Table 7 below.
[0092] Table 7
[0093] In some embodiments, F1 is methionine (M). In some embodiments, F2 is an amino acid selected from the group consisting of G, R, and L. In some embodiments, F3 is an amino acid selected from the group consisting of L, A, and R. In some embodiments, F4 is leucine (L). In some embodiments, F5 is leucine (L). In some embodiments, F6 is an amino acid selected from the group consisting of L and A. In some embodiments, F7 is leucine (L). In some embodiments, F8 is an amino acid selected from the group consisting of L, I, and V. In some embodiments, F9 is leucine (L). In some embodiments, F... 10 It is leucine (L). In some implementations, F 11 It is leucine (L). In some implementations, F 12 These are amino acids selected from the group consisting of L and S. In some embodiments, F... 13 It is leucine (L). In some implementations, F 14 These are amino acids selected from the group consisting of L and V. In some implementations, F... 15 These are amino acids selected from the group consisting of S, P, and G. In some implementations, F... 16 These are amino acids selected from the group consisting of L, P, and G. In some implementations, F... 17 These are amino acids selected from the group consisting of A, V, T, G, and S. In some implementations, F... 18 These are amino acids selected from the group consisting of L and S. In some embodiments, F... 19 These are amino acids selected from the group consisting of A, G, and S.
[0094] It should be understood that variants of SEQ ID NO: 6 do not need to include a substitution at every position providing the substitution, but only one substitution within the scope of Formula VI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 2 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 3 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 6 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 8 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 12 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 14 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and the substitution at position 15 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and a substitution at position 16 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and a substitution at position 17 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and a substitution at position 18 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and a substitution at position 19 of SEQ ID NO: 6. Variations may also comprise any number of substitutions provided by Formula VI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and a substitution at at least one position corresponding to position 2, 3, 6, 8, 12, 14, 15, 16, 17, 18, or 19 of SEQ ID NO: 6. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 6 and substitutions at more than one position corresponding to positions 2, 3, 6, 8, 12, 14, 15, 16, 17, 18 or 19 of SEQ ID NO: 6.
[0095] Variants of SEQ ID NO: 7 (Formula VII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: G1-G2-G3-G4-G5-G6-G7-G8-G9-G 10 -G 11 -G 12 -G 13 -G 14 -G 15 -G 16 -G 17 -G 18 -G 19 -G 20 (Equation VII); Where G1–G 20 The identities of each of them are provided in Table 8 below.
[0096] Table 8
[0097] In some embodiments, G1 is methionine (M). In some embodiments, G2 is an amino acid selected from the group consisting of L, A, and R. In some embodiments, G3 is an amino acid selected from the group consisting of R, P, L, and S. In some embodiments, G4 is an amino acid selected from the group consisting of L, A, and P. In some embodiments, G5 is an amino acid selected from the group consisting of L and P. In some embodiments, G6 is leucine (L). In some embodiments, G7 is an amino acid selected from the group consisting of L, V, and P. In some embodiments, G8 is an amino acid selected from the group consisting of L, A, and V. In some embodiments, G9 is an amino acid selected from the group consisting of L and V. In some embodiments, G... 10 It is leucine (L). In some implementations, G 11 These are amino acids selected from the group consisting of L and V. In some implementations, G... 12 These are amino acids selected from the group consisting of L and A. In some implementations, G... 13 It is leucine (L). In some implementations, G 14 These are amino acids selected from the group consisting of L and A. In some implementations, G... 15 These are amino acids selected from the group consisting of L, V, and A. In some implementations, G... 16 These are amino acids selected from the group consisting of L, A, P, S, and G. In some embodiments, G... 17 These are amino acids selected from the group consisting of L, P, G, and R. In some embodiments, G... 18 These are amino acids selected from the group consisting of V, A, and L. In some implementations, G... 19 These are amino acids selected from the group consisting of R, L, A, and T. In some implementations, G... 20 These are amino acids selected from the group consisting of A, G, and S.
[0098] It should be understood that variations of SEQ ID NO: 7 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula VII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 2 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 3 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 4 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 5 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 7 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 8 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and the substitution at position 9 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 11 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 12 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 14 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 15 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 16 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 17 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 18 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 19 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at position 20 of SEQ ID NO: 7. Variants may also comprise any number of substitutions provided by Formula VII.Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 7, 8, 9, 11, 12, 14, 15, 16, 17, 18, 19, or 20 of SEQ ID NO: 7. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 7 and a substitution at more than one position corresponding to position 2, 3, 4, 5, 7, 8, 9, 11, 12, 14, 15, 16, 17, 18, 19, or 20 of SEQ ID NO: 7.
[0099] Variants of SEQ ID NO: 8 (Formula VIII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: H1-H2-H3-H4-H5-H6-H7-H8-H9-H 10 -H 11 -H 12 -H 13 -H 14 -H 15 -H 16 -H 17 -H 18 -H 19 -H 20 -H 21 (Formula VIII); Where H1–H 21 The identities of each of them are provided in Table 9 below.
[0100] Table 9
[0101] In some embodiments, H1 is methionine (M). In some embodiments, H2 is an amino acid selected from the group consisting of G, T, and S. In some embodiments, H3 is an amino acid selected from the group consisting of M, L, P, and T. In some embodiments, H4 is an amino acid selected from the group consisting of R and H. In some embodiments, H5 is an amino acid selected from the group consisting of L and T. In some embodiments, H6 is an amino acid selected from the group consisting of L and T. In some embodiments, H7 is an amino acid selected from the group consisting of L, C, and M. In some embodiments, H8 is an amino acid selected from the group consisting of L, T, and C. In some embodiments, H9 is an amino acid selected from the group consisting of L, A, V, and T. In some embodiments, H... 10 These are amino acids selected from the group consisting of L, A, and V. In some embodiments, H... 11 These are amino acids selected from the group consisting of L, T, and V. In some implementations, H...12 These are amino acids selected from the group consisting of L and C. In some embodiments, H... 13 These are amino acids selected from the group consisting of L and T. In some embodiments, H... 14 These are amino acids selected from the group consisting of L and S. In some embodiments, H... 15 These are amino acids selected from the group consisting of L and G. In some embodiments, H... 16 These are amino acids selected from the group consisting of A, L, and I. In some implementations, H... 17 These are amino acids selected from the group consisting of P and G. In some implementations, H... 18 These are amino acids selected from the group consisting of P and L. In some embodiments, H... 19 These are amino acids selected from the group consisting of V, A, and I. In some embodiments, H... 20 It is leucine (L). In some implementations, H... 21 It is an amino acid group selected from A, T and G.
[0102] It should be understood that variants of SEQ ID NO: 8 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula VIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 2 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 3 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 4 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 5 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 6 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 7 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and the substitution at position 8 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 9 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 10 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 11 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 12 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 13 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 14 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 15 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 16 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 17 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 18 of SEQ ID NO: 8.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 19 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at position 21 of SEQ ID NO: 8. Variants may also comprise any number of substitutions provided by Formula VIII. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 21 of SEQ ID NO: 8. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 8 and a substitution at more than one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 21 of SEQ ID NO: 8.
[0103] Variants of SEQ ID NO: 9 (Formula IX) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: I1-I2-I3-I4-I5-I6-I7-I8-I9-I 10 -I 11 -I 12 -I 13 -I 14 -I 15 -I 16 -I 17 -I 18 -I 19 -I 20 -I 21 -I 22 (Formula IX); Where I1–I 22 The identities of each of them are provided in Table 10 below.
[0104] Table 10
[0105] In some embodiments, I1 is methionine (M). In some embodiments, I2 is an amino acid selected from the group consisting of A, L, and G. In some embodiments, I3 is an amino acid selected from the group consisting of L, S, and G. In some embodiments, I4 is an amino acid selected from the group consisting of L, P, and G. In some embodiments, I5 is an amino acid selected from the group consisting of R, A, and L. In some embodiments, I6 is an amino acid selected from the group consisting of L and A. In some embodiments, I7 is an amino acid selected from the group consisting of L and P. In some embodiments, I8 is an amino acid selected from the group consisting of L and A. In some embodiments, I9 is an amino acid selected from the group consisting of L and F. In some embodiments, I... 10 These are amino acids selected from the group consisting of L and V. In some embodiments, I 11 These are amino acids selected from the group consisting of L, V, and A. In some implementations, I... 12 These are amino acids selected from the group consisting of L and V. In some embodiments, I... 13 These are amino acids selected from the group consisting of L and F. In some implementations, I... 14 These are amino acids selected from the group consisting of L and A. In some embodiments, I... 15 These are amino acids selected from the group consisting of L, F, and S. In some embodiments, I... 16 It is leucine (L). In some implementations, I 17 These are amino acids selected from the group consisting of L, A, V, and P. In some implementations, I... 18 These are amino acids selected from the group consisting of G, L, A, and P. In some implementations, I... 19 These are amino acids selected from the group consisting of L, G, and V. In some implementations, I... 20 These are amino acids selected from the group consisting of V, A, G, S, and L. In some embodiments, I... 21 These are amino acids selected from the group consisting of L, A, and G. In some implementations, I 22 These are amino acids selected from the group consisting of A, G, and S.
[0106] It should be understood that variants of SEQ ID NO: 9 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula IX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 2 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 3 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 4 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 5 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 6 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 7 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and the substitution at position 8 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 9 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 10 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 11 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 12 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 13 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 14 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 15 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 17 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 18 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 19 of SEQ ID NO: 9.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 20 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 21 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at position 22 of SEQ ID NO: 9. Variants may also comprise any number of substitutions provided by Formula IX. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 21, or 22 of SEQ ID NO: 9. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 9 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 21 or 22 of SEQ ID NO: 9.
[0107] Variants of SEQ ID NO: 10 (Formula X) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: J1-J2-J3-J4-J5-J6-J7-J8-J9-J 10 -J 11 -J 12 -J 13 -J 14 -J 15 -J 16 -J 17 -J 18 -J 19 -J 20 -J 21 -J 22 -J 23 (Formula X); Where J1–J 23 The identities of each of them are provided in Table 11 below.
[0108] Table 11
[0109] In some embodiments, J1 is methionine (M). In some embodiments, J2 is an amino acid selected from the group consisting of A, G, and T. In some embodiments, J3 is an amino acid selected from the group consisting of A, L, P, and G. In some embodiments, J4 is an amino acid selected from the group consisting of L and A. In some embodiments, J5 is an amino acid selected from the group consisting of L and P. In some embodiments, J6 is an amino acid selected from the group consisting of L and A. In some embodiments, J7 is an amino acid selected from the group consisting of L and P. In some embodiments, J8 is leucine (L). In some embodiments, J9 is leucine (L). In some embodiments, J... 10 These are amino acids selected from the group consisting of L and A. In some embodiments, J... 11 These are amino acids selected from the group consisting of L and A. In some embodiments, J... 12 It is leucine (L). In some implementations, J 13 These are amino acids selected from the group consisting of L, A, and V. In some implementations, J... 14 It is leucine (L). In some implementations, J 15 These are amino acids selected from the group consisting of L and A. In some embodiments, J... 16 These are amino acids selected from the group consisting of L and A. In some embodiments, J... 17 These are amino acids selected from the group consisting of L and V. In some embodiments, J 18 These are amino acids selected from the group consisting of L, V, P, and F. In some implementations, J... 19 These are amino acids selected from the group consisting of S, L, and G. In some implementations, J... 20 These are amino acids selected from the group consisting of L, G, and P. In some implementations, J... 21 These are amino acids selected from the group consisting of V, A, and S. In some implementations, J... 22 These are amino acids selected from the group consisting of G and Q. In some implementations, J... 23 These are amino acids selected from the group consisting of A, G, and S.
[0110] It should be understood that variations of SEQ ID NO: 10 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula X. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 2 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 3 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 4 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 5 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 6 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 7 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and the substitution at position 10 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 11 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 13 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 15 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 16 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 17 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 18 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 19 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 20 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 21 of SEQ ID NO: 10.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 22 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at position 23 of SEQ ID NO: 10. Variants may also comprise any number of substitutions provided by Formula X. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 10, 11, 13, 15, 16, 17, 18, 19, 20, 21, 22, or 23 of SEQ ID NO: 10. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 10 and a substitution at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 10, 11, 13, 15, 16, 17, 18, 19, 20, 21, 22, or 23 of SEQ ID NO: 10.
[0111] Variants of SEQ ID NO: 11 (Formula XI) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: K1-K2-K3-K4-K5-K6-K7-K8-K9-K 10 -K 11 -K 12 -K 13 -K 14 -K 15 -K 16 -K 17 -K 18 -K 19 -K 20 -K 21 -K 22 -K 23 -K 24 (Formula XI); Where K1–K 24 The identities of each of them are provided in Table 12 below.
[0112] Table 12
[0113] In some embodiments, K1 is methionine (M). In some embodiments, K2 is an amino acid selected from the group consisting of A, K, and P. In some embodiments, K3 is an amino acid selected from the group consisting of R, A, and L. In some embodiments, K4 is an amino acid selected from the group consisting of P, L, and A. In some embodiments, K5 is an amino acid selected from the group consisting of A, G, P, and S. In some embodiments, K6 is an amino acid selected from the group consisting of L, A, P, and R. In some embodiments, K7 is an amino acid selected from the group consisting of L, R, and S. In some embodiments, K8 is an amino acid selected from the group consisting of L and A. In some embodiments, K9 is an amino acid selected from the group consisting of L and A. In some embodiments, K... 10 These are amino acids selected from the group consisting of L and A. In some implementations, K... 11 These are amino acids selected from the group consisting of L and V. In some implementations, K... 12 These are amino acids selected from the group consisting of L and V. In some implementations, K... 13 These are amino acids selected from the group consisting of L and A. In some implementations, K... 14 These are amino acids selected from the group consisting of L, V, and S. In some implementations, K... 15 It is leucine (L). In some implementations, K 16 These are amino acids selected from the group consisting of L and A. In some implementations, K... 17 These are amino acids selected from the group consisting of L and A. In some implementations, K... 18 These are amino acids selected from the group consisting of L and A. In some implementations, K... 19 These are amino acids selected from the group consisting of L, V, and A. In some implementations, K... 20 These are amino acids selected from the group consisting of P, A, T, and S. In some implementations, K... 21 These are amino acids selected from the group consisting of G, L, and P. In some implementations, K... 22 These are amino acids selected from the group consisting of A, V, S, G, and T. In some implementations, K... 23 These are amino acids selected from the group consisting of L, R, A, and G. In some implementations, K... 24 These are amino acids selected from the group consisting of A, G, and S.
[0114] It should be understood that variations of SEQ ID NO: 11 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 2 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 3 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 4 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 5 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 6 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 7 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and the substitution at position 8 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 9 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 10 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 11 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 12 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 13 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 14 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 16 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 17 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 18 of SEQ ID NO: 11.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 19 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 20 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 21 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 22 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 23 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at position 24 of SEQ ID NO: 11. Variations may also comprise any number of substitutions provided by Formula XI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, or 24 of SEQ ID NO: 11. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 11 and a substitution at more than one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, or 24 of SEQ ID NO: 11.
[0115] Variants of SEQ ID NO: 12 (Formula XII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: L1-L2-L3-L4-L5-L6-L7-L8-L9-L 10 -L 11 -L 12 -L 13 -L 14 -L 15 -L 16 -L 17 -L 18 -L 19 -L 20 -L 21 -L 22 -L 23 -L 24 -L 25 (Formula XII); Where L1–L 25 The identities of each of them are provided in Table 13 below.
[0116] Table 13
[0117] In some embodiments, L1 is methionine (M). In some embodiments, L2 is an amino acid selected from the group consisting of A and G. In some embodiments, L3 is an amino acid selected from the group consisting of A and P. In some embodiments, L4 is an amino acid selected from the group consisting of A, P, and L. In some embodiments, L5 is an amino acid selected from the group consisting of A, P, R, and L. In some embodiments, L6 is an amino acid selected from the group consisting of L, P, A, and G. In some embodiments, L7 is an amino acid selected from the group consisting of L, A, and P. In some embodiments, L8 is an amino acid selected from the group consisting of L, A, and R. In some embodiments, L9 is an amino acid selected from the group consisting of L and P. In some embodiments, L... 10 It is leucine (L). In some implementations, L... 11 It is leucine (L). In some implementations, L... 12 It is leucine (L). In some implementations, L... 13 These are amino acids selected from the group consisting of L and V. In some embodiments, L... 14 These are amino acids selected from the group consisting of L and A. In some embodiments, L... 15 It is leucine (L). In some implementations, L... 16 These are amino acids selected from the group consisting of L and A. In some embodiments, L... 17 These are amino acids selected from the group consisting of L and A. In some embodiments, L... 18 These are amino acids selected from the group consisting of L, A, and G. In some embodiments, L... 19 It is leucine (L). In some implementations, L... 20 These are amino acids selected from the group consisting of L, S, and P. In some embodiments, L... 21 These are amino acids selected from the group consisting of P, G, S, A, and Q. In some implementations, L... 22 These are amino acids selected from the group consisting of G, L, and P. In some implementations, L... 23 These are amino acids selected from the group consisting of A, S, and V. In some implementations, L... 24 These are amino acids selected from the group consisting of R, S, and L. In some embodiments, L... 25 These are amino acids selected from the group consisting of A, G, and S.
[0118] It should be understood that variants of SEQ ID NO: 12 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 2 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 3 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 4 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 5 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 6 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 7 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and the substitution at position 8 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 9 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 13 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 14 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 16 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 17 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 18 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 20 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 21 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 22 of SEQ ID NO: 12.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 23 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 24 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at position 25 of SEQ ID NO: 12. Variants may also comprise any number of substitutions provided by Formula XII. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 13, 14, 16, 17, 18, 20, 21, 22, 23, 24, or 25 of SEQ ID NO: 12. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 12 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 13, 14, 16, 17, 18, 20, 21, 22, 23, 24 or 25 of SEQ ID NO: 12.
[0119] Variants of SEQ ID NO: 13 (Formula XIII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: M1-M2-M3-M4-M5-M6-M7-M8-M9-M 10 -M 11 -M 12 -M 13 -M 14 -M 15 -M 16 -M 17 -M 18 -M 19 -M 20 -M 21 -M 22 -M 23 -M 24 -M 25 -M 26 (Formula XIII); Where M1–M 26 The identities of each of them are provided in Table 14 below.
[0120] Table 14
[0121] In some embodiments, M1 is methionine (M). In some embodiments, M2 is an amino acid selected from the group consisting of A and G. In some embodiments, M3 is an amino acid selected from the group consisting of P, A, S, and L. In some embodiments, M4 is an amino acid selected from the group consisting of P, R, L, A, and G. In some embodiments, M5 is an amino acid selected from the group consisting of G, S, and P. In some embodiments, M6 is an amino acid selected from the group consisting of L, A, and R. In some embodiments, M7 is an amino acid selected from the group consisting of L, A, S, and R. In some embodiments, M8 is an amino acid selected from the group consisting of L, A, G, and P. In some embodiments, M9 is an amino acid selected from the group consisting of L and P. In some embodiments, M... 10 These are amino acids selected from the group consisting of L, R, and P. In some embodiments, M... 11 These are amino acids selected from the group consisting of L and A. In some embodiments, M... 12 These are amino acids selected from the group consisting of L and C. In some embodiments, M... 13 It is leucine (L). In some implementations, M 14 These are amino acids selected from the group consisting of L and V. In some embodiments, M... 15 These are amino acids selected from the group consisting of L and A. In some embodiments, M... 16 These are amino acids selected from the group consisting of L, V, and I. In some embodiments, M... 17 It is leucine (L). In some implementations, M 18 These are amino acids selected from the group consisting of L and A. In some embodiments, M... 19 These are amino acids selected from the group consisting of L, V, and A. In some embodiments, M... 20 It is leucine (L). In some implementations, M 21 These are amino acids selected from the group consisting of L, A, and V. In some embodiments, M... 22 These are amino acids selected from the group consisting of G, P, Q, and S. In some implementations, M... 23 These are amino acids selected from the group consisting of L, V, and P. In some embodiments, M... 24 These are amino acids selected from the group consisting of A, S, V, and T. In some implementations, M... 25 These are amino acids selected from the group consisting of A, L, and S. In some embodiments, M... 26 These are amino acids selected from the group consisting of A, G, and S.
[0122] It should be understood that variants of SEQ ID NO: 13 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 2 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 3 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 4 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 5 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 6 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 7 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and the substitution at position 8 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 9 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 10 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 11 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 12 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 14 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 15 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 16 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 18 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 19 of SEQ ID NO: 13.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 21 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 22 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 23 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 24 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 25 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and a substitution at position 26 of SEQ ID NO: 13. Variations may also comprise any number of substitutions provided by Formula XIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and substitutions at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 18, 19, 21, 22, 23, 24, 25, or 26 of SEQ ID NO: 13. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 13 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 18, 19, 21, 22, 23, 24, 25, or 26 of SEQ ID NO: 13.
[0123] Variants of SEQ ID NO: 14 (Formula XIV) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: N1-N2-N3-N4-N5-N6-N7-N8-N9-N 10 -N 11 -N 12 -N 13 -N 14 -N 15 -N 16 -N 17 -N 18 -N 19 -N 20 -N 21 -N 22 -N 23 -N 24 -N 25 -N 26-N 27 (Form XIV); Where N1–N 27 The identities of each of them are provided in Table 15 below.
[0124] Table 15
[0125] In some embodiments, N1 is methionine (M). In some embodiments, N2 is an amino acid selected from the group consisting of A, G, and R. In some embodiments, N3 is an amino acid selected from the group consisting of P and G. In some embodiments, N4 is an amino acid selected from the group consisting of A, R, G, and L. In some embodiments, N5 is an amino acid selected from the group consisting of G, R, P, and L. In some embodiments, N6 is an amino acid selected from the group consisting of L, A, P, and R. In some embodiments, N7 is an amino acid selected from the group consisting of L, R, and A. In some embodiments, N8 is an amino acid selected from the group consisting of P, R, A, and G. In some embodiments, N9 is an amino acid selected from the group consisting of L, P, and A. In some embodiments, N... 10 These are amino acids selected from the group consisting of L, A, and P. In some implementations, N... 11 These are amino acids selected from the group consisting of L, R, A, and P. In some implementations, N... 12 These are amino acids selected from the group consisting of L and A. In some implementations, N... 13 These are amino acids selected from the group consisting of L and V. In some implementations, N... 14 These are amino acids selected from the group consisting of L, A, and V. In some implementations, N... 15 These are amino acids selected from the group consisting of L and A. In some implementations, N... 16 These are amino acids selected from the group consisting of L, A, and V. In some implementations, N... 17 These are amino acids selected from the group consisting of L, V, and A. In some implementations, N... 18 It is leucine (L). In some implementations, N... 19 These are amino acids selected from the group consisting of L, S, and A. In some implementations, N... 20 These are amino acids selected from the group consisting of L and S. In some implementations, N... 21 These are amino acids selected from the group consisting of L and F. In some implementations, N... 22 These are amino acids selected from the group consisting of L and V. In some implementations, N... 23 These are amino acids selected from the group consisting of G, S, and P. In some implementations, N... 24These are amino acids selected from the group consisting of G, L, A, and P. In some implementations, N... 25 These are amino acids selected from the group consisting of V and A. In some implementations, N... 26 These are amino acids selected from the group consisting of S, L, R, and A. In some implementations, N... 27 It is an amino acid group selected from A and G.
[0126] It should be understood that variants of SEQ ID NO: 14 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XIV. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 2 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 3 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 4 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 5 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 6 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 7 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and the substitution at position 8 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 9 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 10 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 11 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 12 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 13 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 14 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 15 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 16 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 17 of SEQ ID NO: 14.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 19 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 20 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 21 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 22 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 23 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 24 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 25 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 26 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at position 27 of SEQ ID NO: 14. Variants may also comprise any number of substitutions provided by Formula XIV. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 23, 24, 25, 26, or 27 of SEQ ID NO: 14. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 14 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 23, 24, 25, 26 or 27 of SEQ ID NO: 14.
[0127] Variants of SEQ ID NO: 15 (Formula XV) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: O1-O2-O3-O4-O5-O6-O7-O8-O9-O 10 -O 11 -O 12 -O 13 -O14 -O 15 -O 16 -O 17 -O 18 -O 19 -O 20 -O 21 -O 22 -O 23 -O 24 -O 25 -O 26 -O 27 -O 28 (Formula XV); Where O1–O 28 The identities of each of them are provided in Table 16 below.
[0128] Table 16
[0129] In some embodiments, O1 is methionine (M). In some embodiments, O2 is an amino acid selected from the group consisting of A and V. In some embodiments, O3 is an amino acid selected from the group consisting of R and A. In some embodiments, O4 is an amino acid selected from the group consisting of L, A, and R. In some embodiments, O5 is an amino acid selected from the group consisting of R, A, and P. In some embodiments, O6 is an amino acid selected from the group consisting of R and S. In some embodiments, O7 is an amino acid selected from the group consisting of A, L, and G. In some embodiments, O8 is an amino acid selected from the group consisting of P, R, and L. In some embodiments, O9 is an amino acid selected from the group consisting of R, L, and G. In some embodiments, O... 10 These are amino acids selected from the group consisting of P and A. In some implementations, O 11 These are amino acids selected from the group consisting of A, R, and T. In some embodiments, O 12 These are amino acids selected from the group consisting of L, R, and G. In some implementations, O 13 These are amino acids selected from the group consisting of L and A. In some implementations, O 14 These are amino acids selected from the group consisting of L and V. In some implementations, O 15 These are amino acids selected from the group consisting of L and A. In some implementations, O 16 It is leucine (L). In some implementations, O 17 It is leucine (L). In some implementations, O 18 These are amino acids selected from the group consisting of L and F. In some implementations, O 19 It is leucine (L). In some implementations, O 20These are amino acids selected from the group consisting of L and A. In some implementations, O 21 These are amino acids selected from the group consisting of L and A. In some implementations, O 22 These are amino acids selected from the group consisting of L and G. In some implementations, O 23 These are amino acids selected from the group consisting of L and P. In some implementations, O 24 These are amino acids selected from the group consisting of S, A, and T. In some implementations, O 25 These are amino acids selected from the group consisting of L and A. In some implementations, O 26 These are amino acids selected from the group consisting of A, V, G, and S. In some implementations, O... 27 These are amino acids selected from the group consisting of S, R, L, A, and G. In some embodiments, O 28 These are amino acids selected from the group consisting of A, G, and S.
[0130] It should be understood that variants of SEQ ID NO: 15 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XV. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 2 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 3 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 4 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 5 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 6 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 7 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and the substitution at position 8 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 9 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 10 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 11 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 12 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 13 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 14 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 15 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 18 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 20 of SEQ ID NO: 15.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 21 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 22 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 23 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 24 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 25 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 26 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 27 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at position 28 of SEQ ID NO: 15. Variants may also comprise any number of substitutions provided by Formula XV. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 18, 20, 21, 22, 23, 24, 25, 26, 27, or 28 of SEQ ID NO: 15. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 15 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 18, 20, 21, 22, 23, 24, 25, 26, 27, or 28 of SEQ ID NO: 15.
[0131] Variants of SEQ ID NO: 16 (Formula XVI) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: P1-P2-P3-P4-P5-P6-P7-P8-P9-P 10 -P 11 -P 12 -P 13 -P 14 -P 15 -P 16 -P 17 -P18 -P 19 -P 20 -P 21 -P 22 -P 23 -P 24 -P 25 -P 26 -P 27 -P 28 -P 29 (Form XVI); Where P1–P 29 The identities of each of them are provided in Table 17 below.
[0132] Table 17
[0133] In some embodiments, P1 is methionine (M). In some embodiments, P2 is an amino acid selected from the group consisting of A and G. In some embodiments, P3 is an amino acid selected from the group consisting of P and A. In some embodiments, P4 is an amino acid selected from the group consisting of P, A, S, and L. In some embodiments, P5 is an amino acid selected from the group consisting of S and R. In some embodiments, P6 is an amino acid selected from the group consisting of R, L, S, and A. In some embodiments, P7 is an amino acid selected from the group consisting of P, G, R, and L. In some embodiments, P8 is an amino acid selected from the group consisting of G and A. In some embodiments, P9 is an amino acid selected from the group consisting of L, R, and G. In some embodiments, P... 10 These are amino acids selected from the group consisting of P, L, R, and G. In some implementations, P... 11 These are amino acids selected from the group consisting of S, L, and G. In some implementations, P... 12 These are amino acids selected from the group consisting of L, R, and P. In some embodiments, P... 13 These are amino acids selected from the group consisting of L and R. In some embodiments, P 14 It is leucine (L). In some implementations, P 15 It is leucine (L). In some implementations, P 16 It is leucine (L). In some implementations, P 17 It is leucine (L). In some implementations, P 18 It is leucine (L). In some implementations, P 19 It is leucine (L). In some implementations, P 20 These are amino acids selected from the group consisting of L and F. In some implementations, P... 21These are amino acids selected from the group consisting of L and V. In some implementations, P 22 These are amino acids selected from the group consisting of L and A. In some implementations, P... 23 These are amino acids selected from the group consisting of L and A. In some implementations, P... 24 These are amino acids selected from the group consisting of L, W, S, P, and A. In some implementations, P... 25 These are amino acids selected from the group consisting of E and P. In some embodiments, P... 26 These are amino acids selected from the group consisting of P, V, G, and L. In some implementations, P... 27 These are amino acids selected from the group consisting of A, G, and V. In some implementations, P... 28 These are amino acids selected from the group consisting of S and L. In some implementations, P... 29 It is an amino acid group selected from A and G.
[0134] It should be understood that variants of SEQ ID NO: 16 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XVI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 2 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 3 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 4 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 5 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 6 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 7 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and the substitution at position 8 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 9 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 10 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 11 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 12 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 13 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 20 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 21 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 22 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 23 of SEQ ID NO: 16.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 24 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 25 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 26 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 27 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 28 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and a substitution at position 29 of SEQ ID NO: 16. Variations may also comprise any number of substitutions provided by Formula XVI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and substitutions at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 of SEQ ID NO: 16. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 16 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 of SEQ ID NO: 16.
[0135] Variants of SEQ ID NO: 17 (Formula XVII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: Q1-Q2-Q3-Q4-Q5-Q6-Q7-Q8-Q9-Q 10 -Q 11 -Q 12 -Q 13 -Q 14 -Q 15 -Q 16 -Q 17 -Q 18 -Q 19 -Q 20 -Q 21 -Q 22 -Q 23 -Q 24 -Q 25 -Q 26-Q 27 -Q 28 -Q 29 -Q 30 (Form XVII); Where Q1–Q 30 The identities of each of them are provided in Table 18 below.
[0136] Table 18
[0137] In some embodiments, Q1 is methionine (M). In some embodiments, Q2 is an amino acid selected from the group consisting of G, A, E, and K. In some embodiments, Q3 is an amino acid selected from the group consisting of R, A, P, and G. In some embodiments, Q4 is an amino acid selected from the group consisting of G, A, P, L, and S. In some embodiments, Q5 is an amino acid selected from the group consisting of A, G, R, and S. In some embodiments, Q6 is an amino acid selected from the group consisting of G, A, and L. In some embodiments, Q7 is an amino acid selected from the group consisting of R and A. In some embodiments, Q8 is an amino acid selected from the group consisting of P, G, and R. In some embodiments, Q9 is an amino acid selected from the group consisting of A, G, L, and P. In some embodiments, Q... 10 It is an amino acid selected from the group consisting of P and R. In some implementations, Q 11 These are amino acids selected from the group consisting of G, A, L, and P. In some implementations, Q... 12 These are amino acids selected from the group consisting of L, G, R, and S. In some implementations, Q... 13 These are amino acids selected from the group consisting of L, P, and R. In some implementations, Q... 14 These are amino acids selected from the group consisting of L, V, R, and W. In some implementations, Q... 15 It is an amino acid selected from the group consisting of L and P. In some implementations, Q 16 These are amino acids selected from the group consisting of L, V, and F. In some implementations, Q... 17 It is an amino acid selected from the group consisting of L and G. In some implementations, Q 18 These are amino acids selected from the group consisting of L, A, and F. In some implementations, Q... 19 It is an amino acid selected from the group consisting of L and V. In some implementations, Q 20 These are amino acids selected from the group consisting of L and F. In some implementations, Q... 21 It is leucine (L). In some implementations, Q... 22 It is an amino acid selected from the group consisting of L and A. In some implementations, Q 23These are amino acids selected from the group consisting of L, S, and A. In some implementations, Q... 24 It is an amino acid selected from the group consisting of L and A. In some implementations, Q 25 These are amino acids selected from the group consisting of L, F, and T. In some implementations, Q... 26 It is an amino acid selected from the group consisting of P and A. In some implementations, Q 27 These are amino acids selected from the group consisting of G, L, and P. In some implementations, Q... 28 These are amino acids selected from the group consisting of A, T, V, and S. In some implementations, Q... 29 These are amino acids selected from the group consisting of L, R, and Q. In some implementations, Q... 30 These are amino acids selected from the group consisting of A, G, and S.
[0138] It should be understood that variants of SEQ ID NO: 17 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XVII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 2 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 3 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 4 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 5 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 6 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 7 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and the substitution at position 8 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 9 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 10 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 11 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 12 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 13 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 14 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 15 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 16 of SEQ ID NO: 17.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 18 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 19 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 20 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 22 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 23 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 24 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 25 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 26 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 27 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 28 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 29 of SEQ ID NO: 17. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at position 30 of SEQ ID NO: 17. Variations may also comprise any number of substitutions provided by Formula XVII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of SEQ ID NO: 17.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 17 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of SEQ ID NO: 17.
[0139] Variants of SEQ ID NO: 18 (Formula XVIII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 (Formula XVIII); Where R1–R 31 The identities of each of them are provided in Table 19 below.
[0140] Table 19
[0141] In some embodiments, R1 is methionine (M). In some embodiments, R2 is an amino acid selected from the group consisting of A, L, and P. In some embodiments, R3 is an amino acid selected from the group consisting of P, S, A, R, V, and G. In some embodiments, R4 is an amino acid selected from the group consisting of G, P, and R. In some embodiments, R5 is an amino acid selected from the group consisting of G and A. In some embodiments, R6 is an amino acid selected from the group consisting of A, P, and T. In some embodiments, R7 is an amino acid selected from the group consisting of P, A, S, L, and R. In some embodiments, R8 is an amino acid selected from the group consisting of L, A, V, and R. In some embodiments, R9 is an amino acid selected from the group consisting of P, A, S, L, and G. In some embodiments, R... 10 These are amino acids selected from the group consisting of A and L. In some implementations, R... 11 These are amino acids selected from the group consisting of L, P, A, and R. In some embodiments, R... 12 These are amino acids selected from the group consisting of R, V, S, and L. In some embodiments, R... 13 These are amino acids selected from the group consisting of L, T, and A. In some implementations, R... 14 These are amino acids selected from the group consisting of R, L, P, and G. In some embodiments, R... 15 These are amino acids selected from the group consisting of L and R. In some embodiments, R... 16 These are amino acids selected from the group consisting of L, P, and W. In some implementations, R... 17 These are amino acids selected from the group consisting of L, V, and A. In some implementations, R... 18 It is leucine (L). In some implementations, R 19 These are amino acids selected from the group consisting of L and A. In some embodiments, R... 20 These are amino acids selected from the group consisting of L and G. In some implementations, R... 21 These are amino acids selected from the group consisting of L and V. In some implementations, R... 22 These are amino acids selected from the group consisting of L, G, V, and F. In some implementations, R... 23 It is leucine (L). In some implementations, R 24 These are amino acids selected from the group consisting of L, S, and G. In some implementations, R... 25 These are amino acids selected from the group consisting of L, A, and W. In some implementations, R... 26 These are amino acids selected from the group consisting of L, P, A, I, and V. In some implementations, R... 27 These are amino acids selected from the group consisting of G, P, and A. In some implementations, R...28 These are amino acids selected from the group consisting of L, P, S, and A. In some implementations, R... 29 These are amino acids selected from the group consisting of A, V, and S. In some implementations, R... 30 These are amino acids selected from the group consisting of Q, L, A, F, and S. In some implementations, R... 31 It is an amino acid group selected from A and G.
[0142] It should be understood that variants of SEQ ID NO: 18 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XVIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 2 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 3 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 4 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 5 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 6 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 7 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and the substitution at position 8 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 9 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 10 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 11 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 12 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 13 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 14 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 15 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 16 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 17 of SEQ ID NO: 18.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 19 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 20 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 21 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 22 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 24 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 25 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 26 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 27 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 28 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 29 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 30 of SEQ ID NO: 18. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at position 31 of SEQ ID NO: 18. Variants may also comprise any number of substitutions provided by Formula XVIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30 or 31 of SEQ ID NO: 18.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 18 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30 or 31 of SEQ ID NO: 18.
[0143] Variants of SEQ ID NO: 19 (Formula XIX) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: S1-S2-S3-S4-S5-S6-S7-S8-S9-S 10 -S 11 -S 12 -S 13 -S 14 -S 15 -S 16 -S 17 -S 18 -S 19 -S 20 -S 21 -S 22 -S 23 -S 24 -S 25 -S 26 -S 27 -S 28 -S 29 -S 30 -S 31 -S 32 (Formula XIX); Where S1–S 32 The identities of each of them are provided in Table 20 below.
[0144] Table 20
[0145] In some embodiments, S1 is methionine (M). In some embodiments, S2 is an amino acid selected from the group consisting of A, G, R, and S. In some embodiments, S3 is an amino acid selected from the group consisting of A, R, W, L, and G. In some embodiments, S4 is an amino acid selected from the group consisting of P, K, G, and L. In some embodiments, S5 is an amino acid selected from the group consisting of R, G, and P. In some embodiments, S6 is an amino acid selected from the group consisting of A, G, R, and S. In some embodiments, S7 is an amino acid selected from the group consisting of L, A, P, and R. In some embodiments, S8 is an amino acid selected from the group consisting of A, R, G, and P. In some embodiments, S9 is an amino acid selected from the group consisting of P and I. In some embodiments, S... 10 These are amino acids selected from the group consisting of G, P, S, and L. In some implementations, S... 11 These are amino acids selected from the group consisting of L, G, S, and P. In some implementations, S... 12 These are amino acids selected from the group consisting of L, P, G, R, and A. In some implementations, S... 13 These are amino acids selected from the group consisting of L, G, and R. In some embodiments, S... 14 These are amino acids selected from the group consisting of L, R, and T. In some implementations, S... 15 These are amino acids selected from the group consisting of L, S, and P. In some embodiments, S... 16 These are amino acids selected from the group consisting of L, A, and P. In some implementations, S... 17 These are amino acids selected from the group consisting of L, A, C, and T. In some implementations, S... 18 These are amino acids selected from the group consisting of L and V. In some implementations, S 19 These are amino acids selected from the group consisting of L, T, and A. In some implementations, S... 20 It is leucine (L). In some implementations, S 21 These are amino acids selected from the group consisting of L and M. In some embodiments, S... 22 It is leucine (L). In some implementations, S 23 These are amino acids selected from the group consisting of L, A, and V. In some implementations, S... 24 These are amino acids selected from the group consisting of L and G. In some implementations, S... 25 These are amino acids selected from the group consisting of L and A. In some implementations, S... 26 These are amino acids selected from the group consisting of L and S. In some embodiments, S... 27 These are amino acids selected from the group consisting of L, A, P, and G. In some implementations, S...28 These are amino acids selected from the group consisting of P, Q, and S. In some implementations, S... 29 These are amino acids selected from the group consisting of G, V, and L. In some implementations, S... 30 These are amino acids selected from the group consisting of A, S, and V. In some embodiments, S... 31 These are amino acids selected from the group consisting of A, E, R, S, and Q. In some embodiments, S... 32 It is an amino acid group selected from A and G.
[0146] It should be understood that variants of SEQ ID NO: 19 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XIX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 2 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 3 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 4 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 5 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 6 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 7 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and the substitution at position 8 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 9 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 10 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 11 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 12 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 13 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 14 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 15 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 16 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 17 of SEQ ID NO: 19.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 18 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 19 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 21 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 23 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 24 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 25 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 26 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 27 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 28 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 29 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 30 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 31 of SEQ ID NO: 19. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at position 32 of SEQ ID NO: 19. Variations may also comprise any number of substitutions provided by Formula XIX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 23, 24, 25, 26, 27, 28, 29, 30, 31 or 32 of SEQ ID NO: 19.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 19 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 23, 24, 25, 26, 27, 28, 29, 30, 31 or 32 of SEQ ID NO: 19.
[0147] Variants of SEQ ID NO: 20 (Formula XX) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: T1-T2-T3-T4-T5-T6-T7-T8-T9-T 10 -T 11 -T 12 -T 13 -T 14 -T 15 -T 16 -T 17 -T 18 -T 19 -T 20 -T 21 -T 22 -T 23 -T 24 -T 25 -T 26 -T 27 -T 28 -T 29 -T 30 -T 31 -T 32 -T 33 (Formula XX); Where T1–T 33 The identities of each of them are provided in Table 21 below.
[0148] Table 21
[0149] In some embodiments, T1 is methionine (M). In some embodiments, T2 is an amino acid selected from the group consisting of A, G, and E. In some embodiments, T3 is an amino acid selected from the group consisting of A, R, and T. In some embodiments, T4 is an amino acid selected from the group consisting of A, E, and G. In some embodiments, T5 is an amino acid selected from the group consisting of G and R. In some embodiments, T6 is an amino acid selected from the group consisting of R, G, and A. In some embodiments, T7 is an amino acid selected from the group consisting of R, A, P, L, and G. In some embodiments, T8 is an amino acid selected from the group consisting of P, S, G, and R. In some embodiments, T9 is an amino acid selected from the group consisting of R, P, and G. In some embodiments, T... 10 These are amino acids selected from the group consisting of L, R, A, G, S, and P. In some implementations, T... 11 These are amino acids selected from the group consisting of G, P, R, and A. In some implementations, T... 12 These are amino acids selected from the group consisting of P, L, A, R, and G. In some implementations, T... 13 These are amino acids selected from the group consisting of P, L, and S. In some implementations, T... 14 These are amino acids selected from the group consisting of L, S, R, and P. In some implementations, T... 15 These are amino acids selected from the group consisting of A, L, S, and R. In some embodiments, T... 16 These are amino acids selected from the group consisting of L, P, G, V, and A. In some implementations, T... 17 It is leucine (L). In some implementations, T 18 These are amino acids selected from the group consisting of L and P. In some implementations, T... 19 These are amino acids selected from the group consisting of L, A, and R. In some embodiments, T 20 These are amino acids selected from the group consisting of L and G. In some implementations, T... 21 These are amino acids selected from the group consisting of L, S, P, and A. In some implementations, T... 22 These are amino acids selected from the group consisting of L, A, and V. In some implementations, T... 23 These are amino acids selected from the group consisting of L, A, and F. In some implementations, T 24 These are amino acids selected from the group consisting of L and V. In some implementations, T... 25 These are amino acids selected from the group consisting of L and V. In some implementations, T... 26 These are amino acids selected from the group consisting of L and A. In some implementations, T 27These are amino acids selected from the group consisting of L and P. In some implementations, T... 28 These are amino acids selected from the group consisting of A, L, and V. In some implementations, T... 29 These are amino acids selected from the group consisting of P, A, and G. In some implementations, T... 30 These are amino acids selected from the group consisting of G, L, S, and R. In some implementations, T... 31 These are amino acids selected from the group consisting of A, V, and G. In some implementations, T... 32 These are amino acids selected from the group consisting of R, S, G, and T. In some embodiments, T... 33 These are amino acids selected from the group consisting of A, G, and S.
[0150] It should be understood that variations of SEQ ID NO: 20 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 2 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 3 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 4 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 5 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 6 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 7 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and the substitution at position 8 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 9 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 10 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 11 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 12 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 13 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 14 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 15 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 16 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 18 of SEQ ID NO: 20.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 19 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 20 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 21 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 22 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 23 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 24 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 25 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 26 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 27 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 28 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 29 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 30 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 31 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 32 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and a substitution at position 33 of SEQ ID NO: 20. Variants may also comprise any number of substitutions provided by formula XX.Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and substitutions at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 of SEQ ID NO: 20. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 20 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 of SEQ ID NO: 20.
[0151] Variants of SEQ ID NO: 21 (Formula XXI) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: U1-U2-U3-U4-U5-U6-U7-U8-U9-U 10 -U 11 -U 12 -U 13 -U 14 -U 15 -U 16 -U 17 -U 18 -U 19 -U 20 -U 21 -U 22 -U 23 -U 24 -U 25 -U 26 -U 27 -U 28 -U 29 -U 30 -U 31 -U 32 -U 33 -U 34 (Formula XXI); Where U1–U 34 The identities of each of them are provided in Table 22 below.
[0152] Table 22
[0153] In some embodiments, U1 is methionine (M). In some embodiments, U2 is an amino acid selected from the group consisting of G, A, E, and V. In some embodiments, U3 is an amino acid selected from the group consisting of P, R, and H. In some embodiments, U4 is an amino acid selected from the group consisting of P, L, S, R, and A. In some embodiments, U5 is an amino acid selected from the group consisting of S, R, and L. In some embodiments, U6 is an amino acid selected from the group consisting of A, R, and P. In some embodiments, U7 is an amino acid selected from the group consisting of P, G, Q, and S. In some embodiments, U8 is an amino acid selected from the group consisting of P, L, G, and A. In some embodiments, U9 is an amino acid selected from the group consisting of H, C, R, P, G, and S. In some embodiments, U... 10 These are amino acids selected from the group consisting of R, G, A, P, and L. In some implementations, U... 11 These are amino acids selected from the group consisting of L, A, R, P, and E. In some embodiments, U... 12 These are amino acids selected from the group consisting of S, P, C, and A. In some implementations, U... 13 These are amino acids selected from the group consisting of P, L, V, I, and S. In some implementations, U... 14 These are amino acids selected from the group consisting of P, L, R, A, and V. In some implementations, U... 15 These are amino acids selected from the group consisting of S, W, P, and A. In some implementations, U... 16 These are amino acids selected from the group consisting of L, Q, and R. In some implementations, U... 17 These are amino acids selected from the group consisting of G, R, P, and A. In some implementations, U... 18 These are amino acids selected from the group consisting of L, M, and S. In some embodiments, U 19 It is leucine (L). In some implementations, U 20 These are amino acids selected from the group consisting of L and F. In some implementations, U 21 These are amino acids selected from the group consisting of L and T. In some embodiments, U 22 These are amino acids selected from the group consisting of L, A, and S. In some implementations, U... 23 These are amino acids selected from the group consisting of L, S, and V. In some implementations, U... 24 These are amino acids selected from the group consisting of L and V. In some implementations, U 25 It is leucine (L). In some implementations, U 26 These are amino acids selected from the group consisting of L, T, and A. In some implementations, U... 27These are amino acids selected from the group consisting of L, A, F, and S. In some implementations, U... 28 These are amino acids selected from the group consisting of L, W, and C. In some implementations, U... 29 These are amino acids selected from the group consisting of L, N, P, and A. In some implementations, U... 30 These are amino acids selected from the group consisting of P, A, and G. In some implementations, U... 31 It is proline (P). In some implementations, U 32 These are amino acids selected from the group consisting of T, A, V, L, and S. In some implementations, U... 33 These are amino acids selected from the group consisting of T, S, A, and L. In some implementations, U 34 It is an amino acid group selected from A and G.
[0154] It should be understood that variations of SEQ ID NO: 21 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 2 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 3 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 4 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 5 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 6 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 7 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and the substitution at position 8 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 9 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 10 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 11 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 12 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 13 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 14 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 15 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 16 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 17 of SEQ ID NO: 21.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 18 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 20 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 21 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 22 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 23 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 24 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 26 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 27 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 28 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 29 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 30 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 32 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 33 of SEQ ID NO: 21. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at position 34 of SEQ ID NO: 21. Variants may also comprise any number of substitutions provided by Formula XXI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 32, 33 or 34 of SEQ ID NO: 21.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 21 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 32, 33, or 34 of SEQ ID NO: 21.
[0155] Variants of SEQ ID NO: 22 (Formula XXII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: V1-V2-V3-V4-V5-V6-V7-V8-V9-V 10 -V 11 -V 12 -V 13 -V 14 -V 15 -V 16 -V 17 -V 18 -V 19 -V 20 -V 21 -V 22 -V 23 -V 24 -V 25 -V 26 -V 27 -V 28 -V 29 -V 30 -V 31 -V 32 -V 33 -V 34 -V 35 (Formula XXII); Where V1–V 35 The identities of each of them are provided in Table 23 below.
[0156] Table 23
[0157] In some embodiments, V1 is methionine (M). In some embodiments, V2 is an amino acid selected from the group consisting of A and G. In some embodiments, V3 is an amino acid selected from the group consisting of R, A, L, P, and S. In some embodiments, V4 is an amino acid selected from the group consisting of P, A, L, and Q. In some embodiments, V5 is an amino acid selected from the group consisting of G, P, and R. In some embodiments, V6 is an amino acid selected from the group consisting of R and A. In some embodiments, V7 is an amino acid selected from the group consisting of P, S, and A. In some embodiments, V8 is an amino acid selected from the group consisting of P, A, and G. In some embodiments, V9 is an amino acid selected from the group consisting of L, P, and R. In some embodiments, V... 10 These are amino acids selected from the group consisting of R, G, and P. In some embodiments, V... 11 These are amino acids selected from the group consisting of R, A, P, and L. In some implementations, V... 12 These are amino acids selected from the group consisting of R and L. In some embodiments, V 13 These are amino acids selected from the group consisting of P and R. In some embodiments, V 14 These are amino acids selected from the group consisting of P, A, and S. In some implementations, V... 15 These are amino acids selected from the group consisting of G, P, and R. In some embodiments, V... 16 These are amino acids selected from the group consisting of L, Q, and A. In some implementations, V... 17 These are amino acids selected from the group consisting of L and R. In some embodiments, V 18 These are amino acids selected from the group consisting of L, P, S, and R. In some implementations, V... 19 These are amino acids selected from the group consisting of L, W, and R. In some embodiments, V 20 It is leucine (L). In some implementations, V 21 These are amino acids selected from the group consisting of L, T, and P. In some implementations, V... 22 These are amino acids selected from the group consisting of L and A. In some implementations, V... 23 These are amino acids selected from the group consisting of L and S. In some embodiments, V... 24 It is leucine (L). In some implementations, V 25 It is leucine (L). In some implementations, V 26 It is leucine (L). In some implementations, V 27 These are amino acids selected from the group consisting of L, F, and W. In some implementations, V... 28 These are amino acids selected from the group consisting of L and W. In some implementations, V...29 It is leucine (L). In some implementations, V 30 These are amino acids selected from the group consisting of L, P, and V. In some embodiments, V... 31 These are amino acids selected from the group consisting of L, P, and S. In some implementations, V... 32 These are amino acids selected from the group consisting of G, P, and L. In some implementations, V... 33 These are amino acids selected from the group consisting of A, V, T, and S. In some implementations, V... 34 These are amino acids selected from the group consisting of H, L, and V. In some embodiments, V... 35 It is an amino acid group selected from A and G.
[0158] It should be understood that variations of SEQ ID NO: 22 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 2 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 3 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 4 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 5 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 6 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 7 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and the substitution at position 8 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 9 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 10 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 11 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 12 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 13 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 14 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 15 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 16 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 17 of SEQ ID NO: 22.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 18 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 19 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 21 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 22 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 23 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 27 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 28 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 30 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 31 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 32 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 33 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 34 of SEQ ID NO: 22. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at position 35 of SEQ ID NO: 22. Variations may also comprise any number of substitutions provided by Formula XXII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 27, 28, 30, 31, 32, 33, 34 or 35 of SEQ ID NO: 22.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 22 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 27, 28, 30, 31, 32, 33, 34 or 35 of SEQ ID NO: 22.
[0159] Variants of SEQ ID NO: 23 (Formula XXIII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: W1-W2-W3-W4-W5-W6-W7-W8-W9-W 10 -W 11 -W 12 -W 13 -W 14 -W 15 -W 16 -W 17 -W 18 -W 19 -W 20 -W 21 -W 22 -W 23 -W 24 -W 25 -W 26 -W 27 -W 28 -W 29 -W 30 -W 31 -W 32 -W 33 -W 34 -W 35 -W 36 (Formula XXIII); Where W1–W 36 The identities of each of them are provided in Table 24 below.
[0160] Table 24
[0161] In some embodiments, W1 is methionine (M). In some embodiments, W2 is an amino acid selected from the group consisting of A, E, and R. In some embodiments, W3 is an amino acid selected from the group consisting of P, T, L, R, W, and K. In some embodiments, W4 is an amino acid selected from the group consisting of R, P, E, D, A, and H. In some embodiments, W5 is an amino acid selected from the group consisting of G, T, P, W, A, and N. In some embodiments, W6 is an amino acid selected from the group consisting of A, G, I, and M. In some embodiments, W7 is an amino acid selected from the group consisting of R, G, A, and E. In some embodiments, W8 is an amino acid selected from the group consisting of P, G, A, and S. In some embodiments, W9 is an amino acid selected from the group consisting of L, P, Q, G, V, S, and E. In some embodiments, W... 10 These are amino acids selected from the group consisting of R, A, L, V, E, and D. In some implementations, W... 11 These are amino acids selected from the group consisting of G, L, A, D, M, P, and R. In some implementations, W... 12 These are amino acids selected from the group consisting of S, A, D, P, G, and M. In some implementations, W... 13 These are amino acids selected from the group consisting of C, R, D, G, V, A, P, and F. In some implementations, W... 14 These are amino acids selected from the group consisting of G, A, C, S, W, P, and H. In some embodiments, W... 15 These are amino acids selected from the group consisting of T, H, W, R, S, G, A, and I. In some embodiments, W... 16 These are amino acids selected from the group consisting of P, G, R, S, and L. In some implementations, W... 17 These are amino acids selected from the group consisting of A, G, L, and V. In some implementations, W... 18 These are amino acids selected from the group consisting of L, C, K, S, G, Q, and V. In some implementations, W... 19 These are amino acids selected from the group consisting of A, L, G, Q, R, and I. In some implementations, W... 20 These are amino acids selected from the group consisting of S, L, C, Q, R, and A. In some implementations, W... 21 These are amino acids selected from the group consisting of L, A, V, and R. In some implementations, W... 22 These are amino acids selected from the group consisting of V, L, E, F, S, G, and A. In some implementations, W... 23 These are amino acids selected from the group consisting of L, F, Q, T, and C. In some implementations, W... 24 These are amino acids selected from the group consisting of L, S, I, C, and A. In some embodiments, W...25 These are amino acids selected from the group consisting of L, V, I, and F. In some implementations, W... 26 These are amino acids selected from the group consisting of L, F, I, Q, N, and A. In some implementations, W... 27 These are amino acids selected from the group consisting of L, I, S, W, and F. In some embodiments, W... 28 These are amino acids selected from the group consisting of L, I, A, and F. In some implementations, W... 29 These are amino acids selected from the group consisting of L, G, C, I, and F. In some implementations, W... 30 These are amino acids selected from the group consisting of L, W, C, I, T, and S. In some embodiments, W... 31 These are amino acids selected from the group consisting of L, V, C, S, F, P, and G. In some implementations, W... 32 These are amino acids selected from the group consisting of R, Q, T, P, L, and G. In some implementations, W... 33 These are amino acids selected from the group consisting of P, F, D, L, and G. In some implementations, W... 34 These are amino acids selected from the group consisting of S, A, T, C, F, and L. In some implementations, W... 35 These are amino acids selected from the group consisting of R, S, C, E, Q, and G. In some implementations, W... 36 It is an amino acid group selected from A, T, C, P and G.
[0162] It should be understood that variations of SEQ ID NO: 23 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 2 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 3 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 4 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 5 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 6 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 7 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and the substitution at position 8 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 9 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 10 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 11 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 12 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 13 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 14 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 15 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 16 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 17 of SEQ ID NO: 23.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 18 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 19 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 20 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 21 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 22 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 23 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 24 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 25 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 26 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 27 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 28 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 29 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 30 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 31 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 32 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 33 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 34 of SEQ ID NO: 23.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 35 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at position 36 of SEQ ID NO: 23. Variants may also comprise any number of substitutions provided by Formula XXIII. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 of SEQ ID NO: 23. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 23 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 of SEQ ID NO: 23.
[0163] Variants of SEQ ID NO: 24 (Formula XXIV) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36-X 37 (Form XXIV); Where X1–X 37 The identities of each of them are provided in Table 25 below.
[0164] Table 25
[0165] In some embodiments, X1 is methionine (M). In some embodiments, X2 is alanine (A). In some embodiments, X3 is an amino acid selected from the group consisting of A and F. In some embodiments, X4 is an amino acid selected from the group consisting of S and R. In some embodiments, X5 is an amino acid selected from the group consisting of A and L. In some embodiments, X6 is glycine (G). In some embodiments, X7 is an amino acid selected from the group consisting of A, Q, and P. In some embodiments, X8 is an amino acid selected from the group consisting of G and R. In some embodiments, X9 is an amino acid selected from the group consisting of A and G. In some embodiments, X... 10 These are amino acids selected from the group consisting of S, V, and P. In some implementations, X... 11 These are amino acids selected from the group consisting of A, I, S, and C. In some embodiments, X... 12 These are amino acids selected from the group consisting of P, A, Q, and G. In some implementations, X 13 These are amino acids selected from the group consisting of E, A, L, R, and S. In some embodiments, X 14 These are amino acids selected from the group consisting of P, A, N, and R. In some implementations, X... 15 These are amino acids selected from the group consisting of V, E, and L. In some embodiments, X 16 These are amino acids selected from the group consisting of S, A, and P. In some implementations, X... 17 These are amino acids selected from the group consisting of A, R, P, and T. In some implementations, X... 18 These are amino acids selected from the group consisting of R, A, and V. In some embodiments, X 19 These are amino acids selected from the group consisting of L, W, T, and A. In some implementations, X 20 It is an amino acid selected from the group consisting of L and V. In some embodiments, X 21 These are amino acids selected from the group consisting of W, A, and L. In some implementations, X 22 These are amino acids selected from the group consisting of L and S. In some embodiments, X 23 These are amino acids selected from the group consisting of L, V, and R. In some embodiments, X 24These are amino acids selected from the group consisting of L and F. In some implementations, X 25 These are amino acids selected from the group consisting of L and A. In some embodiments, X 26 These are amino acids selected from the group consisting of L, V, and A. In some embodiments, X 27 These are amino acids selected from the group consisting of A and L. In some embodiments, X 28 These are amino acids selected from the group consisting of L and I. In some embodiments, X 29 These are amino acids selected from the group consisting of L, G, and C. In some embodiments, X 30 These are amino acids selected from the group consisting of L and S. In some embodiments, X 31 It is leucine (L). In some implementations, X 32 These are amino acids selected from the group consisting of P, L, and T. In some implementations, X 33 These are amino acids selected from the group consisting of P, A, and V. In some embodiments, X 34 These are amino acids selected from the group consisting of G, S, and L. In some implementations, X 35 These are amino acids selected from the group consisting of V, A, and G. In some implementations, X 36 It is serine (S). In some implementations, X 37 It is an amino acid group selected from A and S.
[0166] It should be understood that variations of SEQ ID NO: 24 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXIV. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 3 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 4 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 5 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 7 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 8 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 9 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and the substitution at position 10 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 11 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 12 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 13 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 14 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 15 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 16 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 17 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 18 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 19 of SEQ ID NO: 24.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 20 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 21 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 22 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 23 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 24 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 25 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 26 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 27 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 28 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 29 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 30 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 32 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 33 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 34 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 35 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at position 37 of SEQ ID NO: 24. Variants may also comprise any number of substitutions provided by Formula XXIV.Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and a substitution at at least one position corresponding to positions 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 33, 34, 35 or 37 of SEQ ID NO: 24. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 24 and substitutions at more than one position corresponding to positions 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 33, 34, 35, or 37 of SEQ ID NO: 24.
[0167] Variants of SEQ ID NO: 25 (Formula XXV) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y 10 -Y 11 -Y 12 -Y 13 -Y 14 -Y 15 -Y 16 -Y 17 -Y 18 -Y 19 -Y 20 -Y 21 -Y 22 -Y 23 -Y 24 -Y 25 -Y 26 -Y 27 -Y 28 -Y 29 -Y 30 -Y 31 -Y 32 -Y 33 -Y 34 -Y 35 -Y 36 -Y 37 -Y 38 (Formula XXV); Where Y1–Y 38 The identities of each of them are provided in Table 26 below.
[0168] Table 26
[0169] In some embodiments, Y1 is methionine (M). In some embodiments, Y2 is an amino acid selected from the group consisting of G, A, E, and S. In some embodiments, Y3 is an amino acid selected from the group consisting of P and L. In some embodiments, Y4 is an amino acid selected from the group consisting of A, G, and P. In some embodiments, Y5 is an amino acid selected from the group consisting of G and A. In some embodiments, Y6 is an amino acid selected from the group consisting of A, T, and D. In some embodiments, Y7 is an amino acid selected from the group consisting of R, A, L, and V. In some embodiments, Y8 is an amino acid selected from the group consisting of R, G, A, and P. In some embodiments, Y9 is an amino acid selected from the group consisting of P and L. In some embodiments, Y... 10 These are amino acids selected from the group consisting of L, G, and R. In some embodiments, Y... 11 These are amino acids selected from the group consisting of G, K, and H. In some implementations, Y... 12 These are amino acids selected from the group consisting of G, S, and F. In some implementations, Y... 13 These are amino acids selected from the group consisting of R and W. In some embodiments, Y 14 These are amino acids selected from the group consisting of R, A, L, and P. In some implementations, Y... 15 These are amino acids selected from the group consisting of P, T, and A. In some implementations, Y... 16 These are amino acids selected from the group consisting of P, G, and L. In some implementations, Y... 17 These are amino acids selected from the group consisting of G, H, and I. In some embodiments, Y... 18 These are amino acids selected from the group consisting of P, M, L, and T. In some implementations, Y... 19 These are amino acids selected from the group consisting of L and P. In some implementations, Y... 20 These are amino acids selected from the group consisting of L and P. In some implementations, Y... 21 It is an amino acid selected from the group consisting of V and R. In some embodiments, Y 22 These are amino acids selected from the group consisting of L and C. In some implementations, Y... 23 These are amino acids selected from the group consisting of L and A. In some embodiments, Y... 24 These are amino acids selected from the group consisting of L, V, and A. In some implementations, Y... 25 These are amino acids selected from the group consisting of L, P, and F. In some implementations, Y... 26 These are amino acids selected from the group consisting of L, I, and A. In some embodiments, Y... 27 These are amino acids selected from the group consisting of L and Y. In some embodiments, Y...28 It is leucine (L). In some implementations, Y 29 These are amino acids selected from the group consisting of L, V, and W. In some implementations, Y... 30 It is leucine (L). In some implementations, Y 31 These are amino acids selected from the group consisting of L, A, and G. In some implementations, Y... 32 These are amino acids selected from the group consisting of L, R, G, and A. In some implementations, Y... 33 These are amino acids selected from the group consisting of S, L, V, P, and A. In some implementations, Y... 34 These are amino acids selected from the group consisting of P, C, R, G, and A. In some embodiments, Y... 35 These are amino acids selected from the group consisting of P and V. In some implementations, Y... 36 It is an amino acid selected from the group consisting of A and G. In some embodiments, Y 37 These are amino acids selected from the group consisting of S, R, Q, and A. In some implementations, Y... 38 It is an amino acid group selected from A and G.
[0170] It should be understood that variations of SEQ ID NO: 25 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXV. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 2 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 3 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 4 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 5 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 6 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 7 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and the substitution at position 8 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 9 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 10 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 11 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 12 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 13 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 14 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 15 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 16 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 17 of SEQ ID NO: 25.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 18 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 19 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 20 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 21 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 22 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 23 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 24 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 25 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 26 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 27 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 29 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 31 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 32 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 33 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 34 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 35 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 36 of SEQ ID NO: 25.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 37 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at position 38 of SEQ ID NO: 25. Variants may also comprise any number of substitutions provided by Formula XXV. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 31, 32, 33, 34, 35, 36, 37, or 38 of SEQ ID NO: 25. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 25 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 31, 32, 33, 34, 35, 36, 37 or 38 of SEQ ID NO: 25.
[0171] Variants of SEQ ID NO: 26 (Formula XXVI) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -Z 20 -Z 21 -Z 22 -Z 23 -Z 24 -Z 25 -Z 26 -Z 27 -Z 28 -Z 29 -Z 30 -Z 31 -Z 32 -Z 33 -Z 34 -Z 35 -Z 36 -Z37 -Z 38 -Z 39 (Form XXVI); Where Z1–Z 39 The identities of each of them are provided in Table 27 below.
[0172] Table 27
[0173] In some embodiments, Z1 is methionine (M). In some embodiments, Z2 is an amino acid selected from the group consisting of A, C, G, L, R, S, and V. In some embodiments, Z3 is an amino acid selected from the group consisting of A, H, Y, K, and L. In some embodiments, Z4 is an amino acid selected from the group consisting of G, A, L, K, and R. In some embodiments, Z5 is an amino acid selected from the group consisting of A, L, R, S, and V. In some embodiments, Z6 is an amino acid selected from the group consisting of P, I, A, S, R, H, and L. In some embodiments, Z7 is an amino acid selected from the group consisting of A, V, E, D, T, and G. In some embodiments, Z8 is an amino acid selected from the group consisting of G, P, A, R, T, and L. In some embodiments, Z9 is an amino acid selected from the group consisting of P, S, K, A, and G. In some embodiments, Z... 10 These are amino acids selected from the group consisting of A, P, S, L, T, and G. In some implementations, Z... 11 These are amino acids selected from the group consisting of R, A, T, H, L, and E. In some embodiments, Z... 12 These are amino acids selected from the group consisting of S, L, K, C, and A. In some implementations, Z... 13 These are amino acids selected from the group consisting of R, V, G, and P. In some implementations, Z... 14 These are amino acids selected from the group consisting of G, A, F, S, and L. In some implementations, Z... 15 These are amino acids selected from the group consisting of A, G, P, M, and V. In some implementations, Z... 16 These are amino acids selected from the group consisting of V, L, R, P, and T. In some implementations, Z... 17 These are amino acids selected from the group consisting of L, A, P, G, R, and V. In some implementations, Z... 18 These are amino acids selected from the group consisting of L, P, A, S, and Y. In some implementations, Z... 19 These are amino acids selected from the group consisting of A, G, V, C, and L. In some implementations, Z... 20 These are amino acids selected from the group consisting of G, F, H, L, C, and A. In some embodiments, Z... 21These are amino acids selected from the group consisting of L, H, C, A, Q, R, and W. In some embodiments, Z... 22 These are amino acids selected from the group consisting of L, R, G, C, M, and E. In some embodiments, Z... 23 These are amino acids selected from the group consisting of L, V, Y, A, and C. In some implementations, Z... 24 These are amino acids selected from the group consisting of L, F, T, I, and V. In some implementations, Z... 25 These are amino acids selected from the group consisting of L, P, S, A, C, and M. In some embodiments, Z... 26 These are amino acids selected from the group consisting of P, V, M, L, and A. In some implementations, Z... 27 These are amino acids selected from the group consisting of L, V, and A. In some implementations, Z... 28 These are amino acids selected from the group consisting of L, G, V, Q, and M. In some implementations, Z... 29 These are amino acids selected from the group consisting of I, L, A, and Q. In some implementations, Z... 30 These are amino acids selected from the group consisting of L, F, V, and T. In some implementations, Z... 31 These are amino acids selected from the group consisting of L, Q, C, A, I, and V. In some implementations, Z... 32 These are amino acids selected from the group consisting of T, F, L, S, and G. In some implementations, Z... 33 These are amino acids selected from the group consisting of S, L, A, and V. In some implementations, Z... 34 These are amino acids selected from the group consisting of A, L, E, S, and G. In some embodiments, Z... 35 These are amino acids selected from the group consisting of P, G, E, and S. In some implementations, Z... 36 These are amino acids selected from the group consisting of G, P, D, W, T, and R. In some implementations, Z... 37 These are amino acids selected from the group consisting of G, A, and S. In some embodiments, Z... 38 These are amino acids selected from the group consisting of S, L, R, and F. In some implementations, Z... 39 It is an amino acid group selected from A and G.
[0174] It should be understood that variations of SEQ ID NO: 26 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXVI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 2 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 3 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 4 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 5 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 6 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 7 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and the substitution at position 8 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 9 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 10 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 11 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 12 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 13 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 14 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 15 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 16 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 17 of SEQ ID NO: 26.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 18 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 19 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 20 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 21 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 22 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 23 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 24 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 25 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 26 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 27 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 28 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 29 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 30 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 31 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 32 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 33 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 34 of SEQ ID NO: 26.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 35 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 36 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 37 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 38 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at position 39 of SEQ ID NO: 26. Variants may also comprise any number of substitutions provided by Formula XXVI. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38 or 39 of SEQ ID NO: 26. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 26 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 of SEQ ID NO: 26.
[0175] Variants of SEQ ID NO: 27 (Formula XXVII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: (AA)1-(AA)2-(AA)3-(AA)4-(AA)5-(AA)6-(AA)7-(AA)8-(AA)9-(AA) 10 -(AA) 11 -(AA) 12 -(AA) 13 -(AA) 14 -(AA) 15 -(AA) 16 -(AA) 17 -(AA) 18 -(AA) 19 -(AA)20 -(AA) 21 -(AA) 22 -(AA) 23 -(AA) 24 -(AA) 25 -(AA) 26 -(AA) 27 -(AA) 28 -(AA) 29 -(AA) 30 -(AA) 31 -(AA) 32 -(AA) 33 -(AA) 34 -(AA) 35 -(AA) 36 -(AA) 37 -(AA) 38 -(AA) 39 -(AA) 40 (Form XXVII); Among them (AA)1–(AA) 40 The identities of each of them are provided in Table 28 below.
[0176] Table 28
[0177] In some embodiments, (AA)1 is methionine (M). In some embodiments, (AA)2 is an amino acid selected from the group consisting of A and W. In some embodiments, (AA)3 is leucine (L). In some embodiments, (AA)4 is an amino acid selected from the group consisting of W, G, P, and Q. In some embodiments, (AA)5 is an amino acid selected from the group consisting of R, Q, and L. In some embodiments, (AA)6 is an amino acid selected from the group consisting of R and S. In some embodiments, (AA)7 is an amino acid selected from the group consisting of P, S, and L. In some embodiments, (AA)8 is an amino acid selected from the group consisting of V, W, and K. In some embodiments, (AA)9 is an amino acid selected from the group consisting of G, P, and T. In some embodiments, (AA) 10 These are amino acids selected from the group consisting of L and G. In some implementations, (AA) 11 These are amino acids selected from the group consisting of E, S, and P. In some implementations, (AA) 12 These are amino acids selected from the group consisting of F, Q, S, and G. In some implementations, (AA) 13 These are amino acids selected from the group consisting of L and G. In some implementations, (AA) 14These are amino acids selected from the group consisting of L, P, and R. In some implementations, (AA) 15 These are amino acids selected from the group consisting of S, H, L, and G. In some implementations, (AA) 16 These are amino acids selected from the group consisting of C, L, and S. In some embodiments, (AA) 17 These are amino acids selected from the group consisting of G, A, and S. In some implementations, (AA) 18 It is tryptophan (W). In some implementations, (AA) 19 These are amino acids selected from the group consisting of R, F, and A. In some implementations, (AA) 20 These are amino acids selected from the group consisting of R, N, and S. In some embodiments, (AA) 21 These are amino acids selected from the group consisting of L, W, Q, and R. In some implementations, (AA) 22 These are amino acids selected from the group consisting of L, M, and S. In some embodiments, (AA) 23 It is leucine (L). In some implementations, (AA) 24 These are amino acids selected from the group consisting of L, G, T, and C. In some implementations, (AA) 25 These are amino acids selected from the group consisting of L and V. In some implementations, (AA) 26 These are amino acids selected from the group consisting of L, F, and P. In some implementations, (AA) 27 These are amino acids selected from the group consisting of V, L, I, A, and G. In some embodiments, (AA) 28 These are amino acids selected from the group consisting of L and A. In some embodiments, (AA) 29 These are amino acids selected from the group consisting of L and A. In some embodiments, (AA) 30 These are amino acids selected from the group consisting of L and I. In some embodiments, (AA) 31 These are amino acids selected from the group consisting of L and A. In some embodiments, (AA) 32 These are amino acids selected from the group consisting of S and L. In some implementations, (AA) 33 These are amino acids selected from the group consisting of L, C, and A. In some embodiments, (AA) 34 These are amino acids selected from the group consisting of R, L, S, and W. In some implementations, (AA) 35 These are amino acids selected from the group consisting of P, L, C, and F. In some implementations, (AA) 36 These are amino acids selected from the group consisting of G, P, and V. In some implementations, (AA) 37These are amino acids selected from the group consisting of G and C. In some embodiments, (AA) 38 These are amino acids selected from the group consisting of S, A, and L. In some implementations, (AA) 39 These are amino acids selected from the group consisting of G, A, and S. In some implementations, (AA) 40 It is an amino acid group selected from A and S.
[0178] It should be understood that variations of SEQ ID NO: 27 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXVII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 2 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 4 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 5 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 6 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 7 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 8 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and the substitution at position 9 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 10 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 11 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 12 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 13 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 14 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 15 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 16 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 17 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 19 of SEQ ID NO: 27.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 20 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 21 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 22 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 24 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 25 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 26 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 27 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 28 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 29 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 30 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 31 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 32 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 33 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 34 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 35 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 36 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 37 of SEQ ID NO: 27.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 38 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 39 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at position 40 of SEQ ID NO: 27. Variants may also comprise any number of substitutions provided by Formula XXVII. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and a substitution at at least one position corresponding to positions 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 of SEQ ID NO: 27. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 27 and substitutions at more than one position corresponding to positions 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 of SEQ ID NO: 27.
[0179] Variants of SEQ ID NO: 28 (Formula XXVIII) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: (BB)1-(BB)2-(BB)3-(BB)4-(BB)5-(BB)6-(BB)7-(BB)8-(BB)9-(BB) 10 -(BB) 11 -(BB) 12 -(BB) 13 -(BB) 14 -(BB) 15 -(BB) 16 -(BB) 17 -(BB) 18 -(BB) 19 -(BB) 20 -(BB) 21 -(BB) 22 -(BB) 23 -(BB) 24 -(BB) 25 -(BB) 26 -(BB)27 -(BB) 28 -(BB) 29 -(BB) 30 -(BB) 31 -(BB) 32 -(BB) 33 -(BB) 34 -(BB) 35 -(BB) 36 -(BB) 37 -(BB) 38 -(BB) 39 -(BB) 40 -(BB) 41 (Formula XXVIII); Where (BB)1–(BB) 41 The identities of each of them are provided in Table 29 below.
[0180] Table 29
[0181] In some embodiments, (BB)1 is methionine (M). In some embodiments, (BB)2 is an amino acid selected from the group consisting of A and G. In some embodiments, (BB)3 is an amino acid selected from the group consisting of S and M. In some embodiments, (BB)4 is an amino acid selected from the group consisting of P and S. In some embodiments, (BB)5 is an amino acid selected from the group consisting of S, R, Q, and P. In some embodiments, (BB)6 is an amino acid selected from the group consisting of S, H, and P. In some embodiments, (BB)7 is an amino acid selected from the group consisting of S, A, and P. In some embodiments, (BB)8 is an amino acid selected from the group consisting of S and L. In some embodiments, (BB)9 is an amino acid selected from the group consisting of W, A, and P. In some embodiments, (BB) 10 These are amino acids selected from the group consisting of L and G. In some implementations, (BB) 11 These are amino acids selected from the group consisting of G, P, and L. In some implementations, (BB) 12 These are amino acids selected from the group consisting of A, V, and G. In some implementations, (BB) 13 These are amino acids selected from the group consisting of R, F, and P. In some implementations, (BB) 14 These are amino acids selected from the group consisting of Q, P, and V. In some implementations, (BB) 15 These are amino acids selected from the group consisting of G, R, and P. In some implementations, (BB) 16These are amino acids selected from the group consisting of P, G, L, and R. In some implementations, (BB) 17 These are amino acids selected from the group consisting of V and P. In some implementations, (BB) 18 These are amino acids selected from the group consisting of L, A, P, G, and Q. In some implementations, (BB) 19 These are amino acids selected from the group consisting of V, G, P, and L. In some implementations, (BB) 20 These are amino acids selected from the group consisting of R, S, P, and M. In some implementations, (BB) 21 These are amino acids selected from the group consisting of A, R, and L. In some implementations, (BB) 22 These are amino acids selected from the group consisting of Q, R, and C. In some implementations, (BB) 23 These are amino acids selected from the group consisting of L and P. In some implementations, (BB) 24 These are amino acids selected from the group consisting of A, L, P, and Q. In some implementations, (BB) 25 These are amino acids selected from the group consisting of L, A, and V. In some implementations, (BB) 26 These are amino acids selected from the group consisting of L and I. In some implementations, (BB) 27 These are amino acids selected from the group consisting of L, A, and P. In some implementations, (BB) 28 These are amino acids selected from the group consisting of L and R. In some implementations, (BB) 29 It is leucine (L). In some implementations, (BB) 30 These are amino acids selected from the group consisting of L, G, and V. In some implementations, (BB) 31 These are amino acids selected from the group consisting of L, P, and A. In some implementations, (BB) 32 These are amino acids selected from the group consisting of L and V. In some implementations, (BB) 33 It is leucine (L). In some implementations, (BB) 34 These are amino acids selected from the group consisting of L, T, G, and P. In some implementations, (BB) 35 These are amino acids selected from the group consisting of A, P, L, and V. In some implementations, (BB) 36 These are amino acids selected from the group consisting of F and L. In some implementations, (BB) 37 These are amino acids selected from the group consisting of P, Q, T, and G. In some implementations, (BB) 38 These are amino acids selected from the group consisting of L, V, G, and T. In some implementations, (BB) 39These are amino acids selected from the group consisting of A, V, and T. In some implementations, (BB) 40 These are amino acids selected from the group consisting of Q, W, L, G, and T. In some implementations, (BB) 41 These are amino acids selected from groups composed of G and A.
[0182] It should be understood that variants of SEQ ID NO: 28 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXVIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 2 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 3 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 4 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 5 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 6 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and the substitution at position 7 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 8 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 9 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 10 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 11 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 12 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 13 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 14 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 15 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 16 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 17 of SEQ ID NO: 28.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 18 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 19 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 20 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 21 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 22 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 23 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 24 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 25 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 26 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 27 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 28 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 30 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 31 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 32 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 34 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 35 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 36 of SEQ ID NO: 28.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 37 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 38 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 39 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 40 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at position 41 of SEQ ID NO: 28. Variants may also comprise any number of substitutions provided by Formula XXVIII. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and a substitution at at least one position corresponding to position 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40 or 41 of SEQ ID NO: 28. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 28 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 30, 31, 32, 34, 35, 36, 37, 38, 39, 40, or 41 of SEQ ID NO: 28.
[0183] Variants of SEQ ID NO: 29 (Formula XXIX) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: (CC)1-(CC)2-(CC)3-(CC)4-(CC)5-(CC)6-(CC)7-(CC)8-(CC)9-(CC) 10 -(CC) 11 -(CC) 12 -(CC) 13 -(CC) 14 -(CC) 15 -(CC) 16 -(CC) 17 -(CC) 18 -(CC) 19 -(CC)20 -(CC) 21 -(CC) 22 -(CC) 23 -(CC) 24 -(CC) 25 -(CC) 26 -(CC) 27 -(CC) 28 -(CC) 29 -(CC) 30 -(CC) 31 -(CC) 32 -(CC) 33 -(CC) 34 -(CC) 35 -(CC) 36 -(CC) 37 -(CC) 38 -(CC) 39 -(CC) 40 -(CC) 41 -(CC) 42 (Formula XXIX); Where (CC)1–(CC) 42 The identities of each of them are provided in Table 30 below.
[0184] Table 30
[0185] In some embodiments, (CC)1 is methionine (M). In some embodiments, (CC)2 is an amino acid selected from the group consisting of A, E, C, G, P, and Q. In some embodiments, (CC)3 is an amino acid selected from the group consisting of R, S, G, N, Q, I, C, and K. In some embodiments, (CC)4 is an amino acid selected from the group consisting of L, R, S, Q, A, C, and E. In some embodiments, (CC)5 is an amino acid selected from the group consisting of G, R, A, P, S, and L. In some embodiments, (CC)6 is an amino acid selected from the group consisting of V, G, W, L, T, and F. In some embodiments, (CC)7 is an amino acid selected from the group consisting of R, E, A, L, S, and I. In some embodiments, (CC)8 is an amino acid selected from the group consisting of R, P, F, W, L, and V. In some embodiments, (CC)9 is an amino acid selected from the group consisting of A, F, R, V, S, and P. In some embodiments, (CC)... 10 These are amino acids selected from the group consisting of P, A, G, T, L, and S. In some embodiments, (CC) 11These are amino acids selected from the group consisting of L, G, A, T, and C. In some embodiments, (CC) 12 These are amino acids selected from the group consisting of P, T, R, A, and E. In some embodiments, (CC) 13 These are amino acids selected from the group consisting of R, G, F, H, E, and V. In some embodiments, (CC) 14 These are amino acids selected from the group consisting of L, G, V, P, S, E, and M. In some embodiments, (CC) 15 These are amino acids selected from the group consisting of P, V, C, R, D, M, and K. In some embodiments, (CC) 16 These are amino acids selected from the group consisting of R, L, G, W, A, and S. In some embodiments, (CC) 17 These are amino acids selected from the group consisting of S, P, T, Q, R, and A. In some implementations, (CC) 18 These are amino acids selected from the group consisting of R, S, N, P, D, and L. In some embodiments, (CC) 19 These are amino acids selected from the group consisting of P, V, D, G, C, and S. In some implementations, (CC) 20 These are amino acids selected from the group consisting of R, G, L, M, K, and H. In some embodiments, (CC) 21 These are amino acids selected from the group consisting of P, R, A, S, and L. In some implementations, (CC) 22 These are amino acids selected from the group consisting of P, R, G, A, W, S, and L. In some embodiments, (CC) 23 These are amino acids selected from the group consisting of L, A, R, P, and Q. In some implementations, (CC) 24 These are amino acids selected from the group consisting of P, L, and A. In some implementations, (CC) 25 These are amino acids selected from the group consisting of L, S, W, and M. In some implementations, (CC) 26 These are amino acids selected from the group consisting of L, R, F, G, and C. In some embodiments, (CC) 27 These are amino acids selected from the group consisting of L, P, and H. In some implementations, (CC) 28 These are amino acids selected from the group consisting of L, W, M, and C. In some implementations, (CC) 29 These are amino acids selected from the group consisting of L, W, A, G, and S. In some embodiments, (CC) 30 These are amino acids selected from the group consisting of L, A, V, and F. In some implementations, (CC) 31 These are amino acids selected from the group consisting of L and W. In some implementations, (CC)32 These are amino acids selected from the group consisting of L, G, V, and T. In some implementations, (CC) 33 These are amino acids selected from the group consisting of L, A, and F. In some implementations, (CC) 34 These are amino acids selected from the group consisting of V, L, S, P, and G. In some implementations, (CC) 35 These are amino acids selected from the group consisting of L, G, and A. In some implementations, (CC) 36 These are amino acids selected from the group consisting of V, N, T, L, F, Q, and G. In some embodiments, (CC) 37 These are amino acids selected from the group consisting of L, P, V, and F. In some implementations, (CC) 38 These are amino acids selected from the group consisting of G, D, H, R, A, S, and P. In some embodiments, (CC) 39 These are amino acids selected from the group consisting of L, P, W, E, S, and G. In some implementations, (CC) 40 These are amino acids selected from the group consisting of A, G, V, and S. In some implementations, (CC) 41 These are amino acids selected from the group consisting of G, A, L, Q, Y, and W. In some implementations, (CC) 42 These are amino acids selected from the group consisting of G, A, and S.
[0186] It should be understood that variations of SEQ ID NO: 29 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XIX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 2 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 3 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 4 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 5 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 6 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 7 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and the substitution at position 8 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 9 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 10 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 11 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 12 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 13 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 14 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 15 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 16 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 17 of SEQ ID NO: 29.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 18 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 19 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 20 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 21 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 22 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 23 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 24 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 25 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 26 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 27 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 28 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 29 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 30 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 31 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 32 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 33 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 34 of SEQ ID NO: 29.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 35 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 36 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 37 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 38 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 39 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 40 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 41 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at position 42 of SEQ ID NO: 29. Variants may also comprise any number of substitutions provided by Formula XXIX. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, or 42 of SEQ ID NO: 29. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 29 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, or 42 of SEQ ID NO: 29.
[0187] Variants of SEQ ID NO: 30 (Formula XXX) In some implementations, the signal peptide comprises an amino acid sequence represented by the following formula: (DD)1-(DD)2-(DD)3-(DD)4-(DD)5-(DD)6-(DD)7-(DD)8-(DD)9-(DD) 10 -(DD) 11 -(DD) 12 -(DD) 13 -(DD) 14 -(DD) 15 -(DD) 16 -(DD) 17 -(DD) 18 -(DD) 19 -(DD) 20 -(DD) 21 -(DD) 22 -(DD) 23 -(DD) 24 -(DD) 25 -(DD) 26 -(DD) 27 -(DD) 28 -(DD) 29 -(DD) 30 -(DD) 31 -(DD) 32 -(DD) 33 -(DD) 34 -(DD) 35 -(DD) 36 -(DD) 37 -(DD)[[ID=5\alpha]] 38 -(DD) 39 -(DD) 40 -(DD) 41 -(DD) 42 -(DD) 43 (Formula XXX); where each of (DD)1–(DD) 43 is provided as in Table 3D below.
[0188] Table 3D
[0189] It should be noted that in the original text, there seems to be a small error in " 38 ", which I temporarily translated as "[[ID=5\alpha]] 38 " for the purpose of translation. You may need to check and correct it according to the actual situation.In some embodiments, (DD)1 is methionine (M). In some embodiments, (DD)2 is an amino acid selected from the group consisting of L, E, and W. In some embodiments, (DD)3 is an amino acid selected from the group consisting of R, G, and I. In some embodiments, (DD)4 is an amino acid selected from the group consisting of A, P, T, and Q. In some embodiments, (DD)5 is an amino acid selected from the group consisting of Q, G, and A. In some embodiments, (DD)6 is an amino acid selected from the group consisting of P, G, M, and L. In some embodiments, (DD)7 is an amino acid selected from the group consisting of L, R, and G. In some embodiments, (DD)8 is an amino acid selected from the group consisting of G and L. In some embodiments, (DD)9 is an amino acid selected from the group consisting of A, W, R, and L. In some embodiments, (DD)... 10 These are amino acids selected from the group consisting of S, G, and L. In some implementations, (DD) 11 These are amino acids selected from the group consisting of P, W, and S. In some implementations, (DD) 12 These are amino acids selected from the group consisting of L, A, and M. In some implementations, (DD) 13 These are amino acids selected from the group consisting of R, L, A, and S. In some implementations, (DD) 14 These are amino acids selected from the group consisting of R, F, A, and I. In some implementations, (DD) 15 These are amino acids selected from the group consisting of R, T, and P. In some implementations, (DD) 16 These are amino acids selected from the group consisting of W, G, and S. In some implementations, (DD) 17 These are amino acids selected from the group consisting of A, V, G, and P. In some implementations, (DD) 18 These are amino acids selected from the group consisting of G, C, and A. In some implementations, (DD) 19 These are amino acids selected from the group consisting of G, V, L, and R. In some implementations, (DD) 20 These are amino acids selected from the group consisting of P and M. In some implementations, (DD) 21 These are amino acids selected from the group consisting of P and L. In some implementations, (DD) 22 These are amino acids selected from the group consisting of S, N, R, and G. In some implementations, (DD) 23 These are amino acids selected from the group consisting of P and S. In some implementations, (DD) 24 These are amino acids selected from the group consisting of L, N, P, and H. In some implementations, (DD) 25These are amino acids selected from the group consisting of L, V, and A. In some implementations, (DD) 26 These are amino acids selected from the group consisting of L, P, and W. In some implementations, (DD) 27 These are amino acids selected from the group consisting of L, S, and I. In some implementations, (DD) 28 These are amino acids selected from the group consisting of L and W. In some implementations, (DD) 29 These are amino acids selected from the group consisting of L and I. In some implementations, (DD) 30 These are amino acids selected from the group consisting of L, T, and A. In some implementations, (DD) 31 These are amino acids selected from the group consisting of L, A, and M. In some implementations, (DD) 32 These are amino acids selected from the group consisting of L and F. In some implementations, (DD) 33 These are amino acids selected from the group consisting of L, A, and Q. In some implementations, (DD) 34 These are amino acids selected from the group consisting of L and I. In some implementations, (DD) 35 These are amino acids selected from the group consisting of W, K, L, and A. In some implementations, (DD) 36 These are amino acids selected from the group consisting of M, F, Q, and V. In some implementations, (DD) 37 These are amino acids selected from the group consisting of L, T, P, and D. In some implementations, (DD) 38 These are amino acids selected from the group consisting of P and L. In some implementations, (DD) 39 These are amino acids selected from the group consisting of P, D, and I. In some implementations, (DD) 40 These are amino acids selected from the group consisting of T, D, P, and A. In some implementations, (DD) 41 These are amino acids selected from the group consisting of V, S, T, and C. In some implementations, (DD) 42 These are amino acids selected from the group consisting of A, Q, W, and E. In some implementations, (DD) 43 It is an amino acid group selected from A and P.
[0190] It should be understood that variations of SEQ ID NO: 30 do not need to include a substitution at every position providing the substitution, but only one substitution within the range of Formula XXX. Therefore, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 2 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 3 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 4 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 5 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 6 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 7 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and the substitution at position 8 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 9 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 10 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 11 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 12 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 13 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 14 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 15 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 16 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 17 of SEQ ID NO: 30.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 18 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 19 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 20 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 21 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 22 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 23 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 24 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 25 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 26 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 27 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 28 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 29 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 30 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 31 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 32 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 33 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 34 of SEQ ID NO: 30.In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 35 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 36 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 37 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 38 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 39 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 40 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 41 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 42 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at position 43 of SEQ ID NO: 30. Variants may also comprise any number of substitutions provided by Formula XXX. Thus, in some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and a substitution at at least one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, or 43 of SEQ ID NO: 30. In some embodiments, the signal peptide comprises the amino acid sequence of SEQ ID NO: 30 and substitutions at more than one position corresponding to positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, ...
Claims
1. A signal peptide comprising an amino acid sequence selected from the group consisting of formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII, XXXIV, XXXV, XXXVI, XXXVII, XXXVIII, or XXXIX; Where equation I is expressed as: A1 - A2 - A3 - A4 - A5 - A6 - A7 - A8 - A9 - A 10 -A 11 -A 12 -A 13 -A 14 (Formula I) And A1-A 14 Their identities are provided in Table 2; Equation II is expressed as: B1 - B2 - B3 - B4 - B5 - B6 - B7 - B8 - B9 - B 10 -B 11 -B 12 -B 13 -B 14 -B 15 (Formula II) And B1-B 15 Their identities are provided in Table 3; Equation III is expressed as: C1-C2-C3-C4-C5-C6-C7-C8-C9-C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 (Formula III) And C1-C 16 Their identities are provided in Table 4; Where equation IV is represented as: D1-D2-D3-D4-D5-D6-D7-D8-D9-D 10 -D 11 -D 12 -D 13 -D 14 -D 15 -D 16 -D 17 (Formula IV) And D1-D 17 Their identities are provided in Table 5; Where V is expressed as: E1-E2-E3-E4-E5-E6-E7-E8-E9-E 10 -E 11 -E 12 -E 13 -E 14 -E 15 -E 16 -E 17 -E 18 (Formula V) And E1-E 18 Their identities are provided in Table 6; Wherein, VI is represented as: F1 - F2 - F3 - F4 - F5 - F6 - F7 - F8 - F9 - F 10 -F 11 -F 12 -F 13 -F 14 -F 15 -F 16 -F 17 -F 18 -F 19 (Formula VI) And F1-F 19 Their identities are provided in Table 7; Equation VII is expressed as: G1 - G2 - G3 - G4 - G5 - G6 - G7 - G8 - G9 - G 10 -G 11 -G 12 -G 13 -G 14 -G 15 -G 16 -G 17 -G 18 -G 19 -G 20 (Formula VII) And G1-G 20 Their identities are provided in Table 8; Where equation VIII is expressed as: H1-H2-H3-H4-H5-H6-H7-H8-H9-H 10 -H 11 -H 12 -H 13 -H 14 -H 15 -H 16 -H 17 -H 18 -H 19 -H 20 -H 21 (Formula VIII) And H1-H 21 Their identities are provided in Table 9; Where IX is represented as: I1 - I2 - I3 - I4 - I5 - I6 - I7 - I8 - I9 - I 10 -I 11 -I 12 -I 13 -I 14 -I 15 -I 16 -I 17 -I 18 -I 19 -I 20 -I 21 -I 22 (Formula IX) And I1-I 22 Their identities are provided in Table 10; Where X is represented as: J1-J2-J3-J4-J5-J6-J7-J8-J9-J 10 -J 11 -J 12 -J 13 -J 14 -J 15 -J 16 -J 17 -J 18 -J 19 -J 20 -J 21 -J 22 -J 23 (Formula X) And J1-J 23 Their identities are provided in Table 11; Where XI is represented as: K1-K2-K3-K4-K5-K6-K7-K8-K9-K 10 -K 11 -K 12 -K 13 -K 14 -K 15 -K 16 -K 17 -K 18 -K 19 -K 20 -K 21 -K 22 -K 23 -K 24 (Formula XI) And K1-K 24 Their identities are provided in Table 12; Where XII is expressed as: L1-L2-L3-L4-L5-L6-L7-L8-L9-L 10 -L 11 -L 12 -L 13 -L 14 -L 15 -L 16 -L 17 -L 18 -L 19 -L 20 -L 21 -L 22 -L 23 -L 24 -L 25 (Formula XII) And L1-L 25 Their identities are provided in Table 13; Where XIII is expressed as: M1 - M2 - M3 - M4 - M5 - M6 - M7 - M8 - M9 - M 10 -M 11 -M 12 -M 13 -M 14 -M 15 -M 16 -M 17 -M 18 -M 19 -M 20 -M 21 -M 22 -M 23 -M 24 -M 25 -M 26 (Formula XIII) And M1-M 26 Their identities are provided in Table 14; Where XIV is expressed as: N1-N2-N3-N4-N5-N6-N7-N8-N9-N 10 -N 11 -N 12 -N 13 -N 14 -N 15 -N 16 -N 17 -N 18 -N 19 -N 20 -N 21 -N 22 -N 23 -N 24 -N 25 -N 26 -N 27 (Formula XIV) And N1-N 27 Their identities are provided in Table 15; Where XV is expressed as: O1 - O2 - O3 - O4 - O5 - O6 - O7 - O8 - O9 - O 10 -O 11 -O 12 -O 13 -O 14 -O 15 -O 16 -O 17 -O 18 -O 19 -O 20 -O 21 -O 22 -O 23 -O 24 -O 25 -O 26 -O 27 -O 28 (Formula XV) And O1-O 28 Their identities are provided in Table 16; Where XVI is expressed as: P1 - P2 - P3 - P4 - P5 - P6 - P7 - P8 - P9 - P 10 -P 11 -P 12 -P 13 -P 14 -P 15 -P 16 -P 17 -P 18 -P 19 -P 20 -P 21 -P 22 -P 23 -P 24 -P 25 -P 26 -P 27 -P 28 -P 29 (Formula XVI) And P1-P 29 Their identities are provided in Table 17; Where XVII is expressed as: Q1-Q2-Q3-Q4-Q5-Q6-Q7-Q8-Q9-Q 10 -Q 11 -Q 12 -Q 13 -Q 14 -Q 15 -Q 16 -Q 17 -Q 18 -Q 19 -Q 20 -Q 21 -Q 22 -Q 23 -Q 24 -Q 25 -Q 26 -Q 27 -Q 28 -Q 29 -Q 30 (Formula XVII) And Q1-Q 30 Their identities are provided in Table 18; Where XVIII is expressed as: R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 (Formula XVIII) And R1-R 31 Their identities are provided in Table 19; Where XIX is represented as: S1-S2-S3-S4-S5-S6-S7-S8-S9-S 10 -S 11 -S 12 -S 13 -S 14 -S 15 -S 16 -S 17 -S 18 -S 19 -S 20 -S 21 -S 22 -S 23 -S 24 -S 25 -S 26 -S 27 -S 28 -S 29 -S 30 -S 31 -S 32 (Formula XIX) And S1-S 32 The identities are provided in Table 20; Where XX is represented as: T1-T2-T3-T4-T5-T6-T7-T8-T9-T 10 -T 11 -T 12 -T 13 -T 14 -T 15 -T 16 -T 17 -T 18 -T 19 -T 20 -T 21 -T 22 -T 23 -T 24 -T<00 And T1-T 33 Their identities are provided in Table 21; Where XXI is represented as: U1-U2-U3-U4-U5-U6-U7-U8-U9-U 10 -U 11 -U 12 -U 13 -U 14 -U 15 -U 16 -U 17 -U 18 -U 19 -U 20 -U 21 -U 22 -U 23 -U 24 -U 25 -U 26 -U 27 -U 28 -U 29 -U 30 -U 31 -U 32 -U 33 -U 34 (Formula XXI) And U1-U 34 Their identities are provided in Table 22; Where equation XXII is represented as: V1-V2-V3-V4-V5-V6-V7-V8-V9-V 10 -V 11 -V 12 -V 13 -V 14 -V 15 -V 16 -V 17 -V 18 -V 19 -V 20 -V 21 -V 22 -V 23 -V 24 -V 25 -V 26 -V 27 -V 28 -V 29 -V 30 -V 31 -V 32 -V 33 -V 34 -V 35 (Formula XXII) And V1-V 35 Their identities are provided in Table 23; Where equation XXIII is represented as: W1 - W2 - W3 - W4 - W5 - W6 - W7 - W8 - W9 - W 10 -W 11 -W 12 -W 13 -W 14 -W 15 -W 16 -W 17 -W 18 -W 19 -W 20 -W 21 -W 22 -W 23 -W 24 -W 25 -W 26 -W 27 -W 28 -W 29 -W 30 -W 31 -W 32 -W 33 -W 34 -W 35 -W 36 (Formula XXIII) And W1-W 36 Their identities are provided in Table 24; Where equation XXIV is represented as: X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36 -X 37 (Formula XXIV) And X1-X 37 Their identities are provided in Table 25; Where XXV is represented as: Y1 - Y2 - Y3 - Y4 - Y5 - Y6 - Y7 - Y8 - Y9 - Y 10 -Y 11 -Y 12 -Y 13 -Y 14 -Y 15 -Y 16 -Y 17 -Y 18 -Y 19 -Y 20 -Y 21 -Y 22 -Y 23 -Y 24 -Y 25 -Y 26 -Y 27 -Y 28 -Y 29 -Y 30 -Y 31 -Y 32 -Y 33 -Y 34 -Y 35 -Y 36 -Y 37 -Y 38 (Formula XXV) And Y1-Y 38 Their identities are provided in Table 26; Wherein, XXVI is represented as: Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -Z 20 -Z 21 -Z 22 -Z 23 -Z 24 -Z 25 -Z 26 -Z 27 -Z 28 -Z 29 -Z 30 -Z 31 -Z 32 -Z 33 -Z 34 -Z 35 -Z 36 -Z 37 -Z 38 -Z 39 (Formula XXVI) And Z1-Z 39 Their identities are provided in Table 27; Where equation XXVII is represented as: (AA)1-(AA)2-(AA)3-(AA)4-(AA)5-(AA)6-(AA)7-(AA)8-(AA)9-(AA) 10 -(AA) 11 -(AA) 12 -(AA) 13 -(AA) 14 -(AA) 15 -(AA) 16 -(AA) 17 -(AA) 18 -(AA) 19 -(AA) 20 -(AA) 21 -(AA) 22 -(AA) 23 -(AA) 24 -(AA) 25 -(AA) 26 -(AA) 27 -(AA) 28 -(AA) 29 -(AA) 30 -(AA) 31 -(AA) 32 -(AA) 33 -(AA) 34 -(AA) 35 -(AA) 36 -(AA) 37 -(AA) 38 -(AA) 39 -(AA) 40 (Formula XXVII) And (AA)1-(AA) 40 Their identities are provided in Table 28; Where equation XXVIII is represented as: (BB)1-(BB)2-(BB)3-(BB)4-(BB)5-(BB)6-(BB)7-(BB)8-(BB)9-(BB) 10 -(BB) 11 -(BB) 12 -(BB) 13 -(BB) 14 -(BB) 15 -(BB) And (BB)1-(BB) 41 Their identities are provided in Table 29; Where XXIX is represented as: (CC)1-(CC)2-(CC)3-(CC)4-(CC)5-(CC)6-(CC)7-(CC)8-(CC)9-(CC) 10 -(CC) 11 -(CC) 12 -(CC) 13 -(CC) 14 -(CC) 15 -(CC) 16 -(CC) 17 -(CC) 18 -(CC) 19 -(CC) 20 -(CC) 21 -(CC) 22 -(CC) 23 -(CC) 24 -(CC) 25 -(CC) 26 -(CC) 27 -(CC) 28 -(CC) 29 -(CC) 30 -(CC) 31 -(CC) 32 -(CC) 33 -(CC) 34 -(CC) 35 -(CC) 36 -(CC) 37 -(CC) 38 -(CC) 39 -(CC) 40 -(CC) 41 -(CC) 42 (Formula XXIX) And (CC)1-(CC) 42 Their identities are provided in Table 30; Where the formula XXX is represented as: (DD)1-(DD)2-(DD)3-(DD)4-(DD)5-(DD)6-(DD)7-(DD)8-(DD)9-(DD) 10 -(DD) 11 -(DD) 12 -(DD) 13 -(DD) 14 -(DD) 15 -(DD) 16 -(DD) 17 -(DD) 18 -(DD) 19 -(DD) 20 -(DD) 21 -(DD) 22 -(DD) 23 -(DD) 24 -(DD) 25 -(DD) 26 -(DD) 27 -(DD) 28 -(DD) 29 -(DD) 30 -(DD) 31 -(DD) 32 -(DD) 33 -(DD) 34 -(DD) 35 -(DD) 36 -(DD) 37 -(DD)<00006 And (DD)1-(DD) 43 Their identities are provided in Table 31; Wherein, XXXI is represented as: (EE)1-(EE)2-(EE)3-(EE)4-(EE)5-(EE)6-(EE)7-(EE)8-(EE)9-(EE) 10 -(EE) 11 -(EE) 12 -(EE) 13 -(EE) 14 -(EE) 15 -(EE) 16 -(EE) 17 -(EE) 18 -(EE) 19 -(EE) 20 -(EE) 21 -(EE) 22 -(EE) 23 -(EE) 24 -(EE) 25 -(EE) 26 -(EE) 27 -(EE) 28 -(EE) 29 -(EE) 30 -(EE) 31 -(EE) 32 -(EE) 33 -(EE) 34 -(EE) 35 -(EE) 36 -(EE) 37 -(EE) 38 -(EE) 39 -(EE) 40 -(EE) 41 -(EE) 42 -(EE) 43 -(EE) 44 (Formula XXXI) And (EE)1-(EE) 44 Their identities are provided in Table 32; Wherein, equation XXXII is represented as: (FF)1-(FF)2-(FF)3-(FF)4-(FF)5-(FF)6-(FF)7-(FF)8-(FF)9-(FF) 10 -(FF) 11 -(FF) 12 -(FF) 13 -(FF) 14 -(FF) 15 -(FF) 16 -(FF) 17 -(FF) 18 -(FF) 19 -(FF) 20 -(FF) 21 -(FF) 22 -(FF) 23 -(FF) 24 -(FF) 25 -(FF) 26 -(FF) 27 -(FF) 28 -(FF) 29 -(FF) 30 -(FF) 31 -(FF) 32 -(FF) 33 -(FF) 34 -(FF) 35 -(FF) 36 -(FF) 37 -(FF) 38 -(FF) 39 -(FF) 40 -(FF) 41 -(FF) 42 -(FF) 43 -(FF) 44 -(FF) 45 (Formula XXXII) And (FF)1-(FF) 45 Their identities are provided in Table 33; Where equation XXXIII is represented as: (GG)1-(GG)2-(GG)3-(GG)4-(GG)5-(GG)6-(GG)7-(GG)8-(GG)9-(GG) 10 -(GG) 11 -(GG) 12 -(GG) 13 -(GG) 14 -(GG) 15 -(GG) 16 -(GG) 17 -(GG) 18 -(GG) 19 -(GG) 20 -(GG) 21 -(GG) 22 -(GG) 23 -(GG) 24 -(GG) 25 -(GG) 26 -(GG) 27 -(GG) 28 -(GG) 29 -(GG) 30 -(GG) 31 -(GG) 32 -(GG) 33 -(GG) 34 -(GG) 35 -(GG) 36 And (GG)1-(GG) 46 Their identities are provided in Table 34; Wherein, XXXIV is represented as: (HH)1-(HH)2-(HH)3-(HH)4-(HH)5-(HH)6-(HH)7-(HH)8-(HH)9-(HH) 10 -(HH) 11 -(HH) 12 -(HH) 13 -(HH) 14 -(HH) 15 -(HH) 16 ... (Formula XXXIV) And (HH)1-(HH) 47 Their identities are provided in Table 35; Wherein, XXXV is represented as: (II)1-(II)2-(II)3-(II)4-(II)5-(II)6-(II)7-(II)8-(II)9-(II) 10 -(II) 11 -(II) 12 -(II) 13 -(II) 14 -(II) 15 -(II) 16 -(II) 17 -(II) 18 -(II) 19 -(II) 20 -(II) 21 -(II) 22 -(II) 23 -(II) 24 -(II) 25 -(II) 26 -(II) 27 -(II) 28 -(II) 29 )-(II) 30 -(II) 31 -(II) 32 -(II) 33 -(II) 34 -(II) 35 -(II) 36 -(II) 37 -(II) 38 -(II) 39 -(II) 40 -(II) 41 -(II) 42 -(II) 43 -(II) 44 -(II) 45 -(II) 46 -(II) 47 -(II) 48 (Formula XXXV) And (II)1-(II) 48 The identities are provided in Table 36; Wherein, XXXVI is represented as: (JJ)1-(JJ)2-(JJ)3-(JJ)4-(JJ)5-(JJ)6-(JJ)7-(JJ)8-(JJ)9-(JJ) 10 -(JJ) 11 -(JJ) 12 -(JJ) 13 -(JJ) 14 -(JJ) 15 -(JJ) 16 -(JJ) 17 -(JJ) 18 -(JJ) 19 -(JJ) 20 -(JJ) 21 -(JJ) 22 -(JJ) 23 -(JJ) 24 -(JJ) 25 -(JJ) 26 -(JJ)<000 And (JJ)1-(JJ) 49 Their identities are provided in Table 37; Where equation XXXVII is represented as: (KK)1-(KK)2-(KK)3-(KK)4-(KK)5-(KK)6-(KK)7-(KK)8-(KK)9-(KK) 10 -(KK) 11 -(KK) 12 -(KK) 13 -(KK) 14 -(KK) 15 -(KK) 16 -(KK) 17 -(KK) 18 -(KK) 19 -(KK) 20 -(KK) 21 -(KK) 22 -(KK) 23 -(KK) 24 -(KK) 25 -(KK) 26 -(KK) 27 -(KK) 28 -(KK) 29 -(KK) 30 -(KK) 31 -(KK) 32 -(KK) 33 -(KK) 34 -(KK) 35 -(KK) 36 -(KK) 37 -(KK) 38 -(KK) 39 -(KK) 40 -(KK) 41 -(KK) 42 -(KK) 43 -(KK) 44 -(KK) 45 -(KK) 46 -(KK)[[ID=7)) 47 -(KK) 48 [[ID=H8]]-(KK) 49 -(KK) 50 -(KK) 51 -(KK) 52 (Formula XXXVII) And (KK)1-(KK) 52 The identities are provided in Table 38; Wherein, equation XXXVIII is represented as: (LL)1-(LL)2-(LL)3-(LL)4-(LL)5-(LL)6-(LL)7-(LL)8-(LL)9-(LL) 10 -(LL) 11 -(LL) 12 -(LL) 13 -(LL) 14 -(LL) 15 -(LL) 16 -(LL) 17 -(LL) 18 -(LL) 19 -(LL) 20 -(LL) 21 -(LL) 22 -(LL) 23 -(LL) 24 -(LL) 25 -(LL) 26 -(LL) 27 -(LL) 28 -(LL) 29 -(LL) 30 -(LL) 31 -(LL) 32 -(LL) 33 -(LL) 34 -(LL) 35 -(LL) 36 -(LL) 37 -(LL) 38 -(LL) 39 -(LL) 40 -(LL) 41 -(LL) 42 -(LL) 43 -(LL) 44 -(LL) 45 -(LL) 46 -(LL) 47 -(LL) 48 -(LL) 49 -(LL) 50 -(LL) 51 -(LL) 52 -(LL) 53 -(LL) 54 -(LL) 55 (Formula XXXVIII) And (LL)1-(LL) 55 The identities are provided in Table 39; and Where the expression XXXIX is represented as: (MM)1-(MM)2-(MM)3-(MM)4-(MM)5-(MM)6-(MM)7-(MM)8-(MM)9-(MM) 10 -(MM) 11 -(MM) 12 -(MM) 13 -(MM) 14 -(MM) 15 -(MM) 16 -(MM) 17 -(MM) 18 -(MM) 19 -(MM) 20 -(MM) 21 -(MM) 22 -(MM) 23 -(MM) 24 -(MM) 25 -(MM) 26 -(MM) 27 -(MM) 28 -(MM) 29 -(MM) 30 -(MM) 31 -(MM) 32 -(MM) 33 -(MM) 34 -(MM) 35 -(MM) 36 -(MM) 37 -(MM) 38 -(MM) 39 -(MM) 40 -(MM) 41 -(MM) 42 -(MM) 43 -(MM) 44 -(MM) 45 -(MM) 46 -(MM) 47 -(MM) 48 -(MM) 49 -(MM) 50 -(MM) 51 -(MM) 52 -(MM) 53 -(MM) 54 -(MM) 55 -(MM) 56 -(MM) 57 -(MM) 58 -(MM) 59 (Formula XXXIX) And (MM)1-(MM) 59 Their identities are provided in Table 40.
2. The signal peptide of claim 1, comprising an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 99, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
3. A signal peptide comprising an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 9, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
4. The signal peptide according to any one of claims 1 to 3, wherein the signal peptide is a novel signal peptide.
5. A subset X1-(Y1) a -Z1 recombinant polypeptide, wherein: X1 is a signal peptide. Y1 is a peptide linker, and Z1 is the payload protein. Where a is an integer selected from 0 and 1.
6. The recombinant polypeptide of claim 5, wherein X1 comprises an amino acid sequence selected from the group consisting of formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, XXVIII, XXIX, XXX, XXXI, XXXII, XXXIII, XXXIV, XXXV, XXXVI, XXXVII, XXXVIII, or XXXIX; Where equation I is expressed as: A1 - A2 - A3 - A4 - A5 - A6 - A7 - A8 - A9 - A 10 -A 11 -A 12 -A 13 -A 14 (Formula I) And A1-A 14 Their identities are provided in Table 2; Equation II is expressed as: B1 - B2 - B3 - B4 - B5 - B6 - B7 - B8 - B9 - B 10 -B 11 -B 12 -B 13 -B 14 -B 15 (Formula II) And B1-B 15 Their identities are provided in Table 3; Equation III is expressed as: C1-C2-C3-C4-C5-C6-C7-C8-C9-C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 (Formula III) And C1-C 16 Their identities are provided in Table 4; Where equation IV is represented as: D1 - D2 - D3 - D4 - D5 - D6 - D7 - D8 - D9 - D 10 -D 11 -D 12 -D 13 -D 14 -D 15 -D 16 -D 17 (Formula IV) And D1-D 17 Their identities are provided in Table 5; Where V is expressed as: E1-E2-E3-E4-E5-E6-E7-E8-E9-E 10 -E 11 -E 12 -E 13 -E 14 -E 15 -E 16 -E 17 -E 18 (Formula V) And E1-E 18 Their identities are provided in Table 6; Wherein, VI is represented as: F1 - F2 - F3 - F4 - F5 - F6 - F7 - F8 - F9 - F 10 -F 11 -F 12 -F 13 -F 14 -F 15 -F 16 -F 17 -F 18 -F 19 (Formula VI) And F1-F 19 Their identities are provided in Table 7; Equation VII is expressed as: G1 - G2 - G3 - G4 - G5 - G6 - G7 - G8 - G9 - G 10 -G 11 -G 12 -G 13 -G 14 -G 15 -G 16 -G 17 -G 18 -G 19 -G 20 (Formula VII) And G1-G 20 Their identities are provided in Table 8; Where equation VIII is expressed as: H1-H2-H3-H4-H5-H6-H7-H8-H9-H 10 -H 11 -H 12 -H 13 -H 14 -H 15 -H 16 -H 17 -H 18 -H 19 -H 20 -H 21 (Formula VIII) And H1-H 21 Their identities are provided in Table 9; Where IX is represented as: I1 - I2 - I3 - I4 - I5 - I6 - I7 - I8 - I9 - I 10 -I 11 -I 12 -I 13 -I 14 -I 15 -I 16 -I 17 -I 18 -I 19 -I 20 -I 21 -I 22 (Formula IX) And I1-I 22 Their identities are provided in Table 10; Where X is represented as: J1-J2-J3-J4-J5-J6-J7-J8-J9-J 10 -J 11 -J 12 -J 13 -J 14 -J 15 -J 16 -J 17 -J 18 -J 19 -J 20 -J 21 -J 22 -J 23 (Formula X) And J1-J 23 Their identities are provided in Table 11; Where XI is represented as: K1-K2-K3-K4-K5-K6-K7-K8-K9-K 10 -K 11 -K 12 -K 13 -K 14 -K 15 -K 16 -K 17 -K 18 -K 19 -K 20 -K 21 -K 22 -K 23 -K 24 (Formula XI) And K1-K 24 Their identities are provided in Table 12; Where XII is expressed as: L1-L2-L3-L4-L5-L6-L7-L8-L9-L 10 -L 11 -L 12 -L 13 -L 14 -L 15 -L 16 -L 17 -L 18 -L 19 -L 20 -L 21 -L 22 -L 23 -L 24 -L 25 (Formula XII) And L1-L 25 Their identities are provided in Table 13; Where XIII is expressed as: M1-M2-M3-M4-M5-M6-M7-M8-M9-M 10 -M 11 -M 12 -M 13 -M 14 -M 15 -M 16 -M 17 -M 18 -M 19 -M 20 -M 21 -M 22 -M 23 -M 24 -M 25 -M 26 (Formula XIII) And M1-M 26 Their identities are provided in Table 14; Where XIV is expressed as: N1-N2-N3-N4-N5-N6-N7-N8-N9-N 10 -N 11 -N 12 -N 13 -N 14 -N 15 -N 16 -N 17 -N 18 -N 19 -N 20 -N 21 -N 22 -N 23 -N 24 -N 25 -N 26 -N 27 (Formula XIV) And N1-N 27 Their identities are provided in Table 15; Where XV is expressed as: O1 - O2 - O3 - O4 - O5 - O6 - O7 - O8 - O9 - O 10 -O 11 -O 12 -O 13 -O 14 -O 15 -O 16 -O 17 -O 18 -O 19 -O 20 -O 21 -O 22 -O 23 -O 24 -O 25 -O 26 -O 27 -O 28 (Formula XV) And O1-O 28 Their identities are provided in Table 16; Where XVI is expressed as: P1 - P2 - P3 - P4 - P5 - P6 - P7 - P8 - P9 - P 10 -P 11 -P 12 -P 13 -P 14 -P 15 -P 16 -P 17 -P 18 -P 19 [[ID=2))-P 20 -P 21 -P 22 -P 23 -P 24 -P 25 -P 26 -P 27 -P 28 -P 29 (Formula XVI) And P1-P 29 Their identities are provided in Table 17; Where XVII is expressed as: Q1-Q2-Q3-Q4-Q5-Q6-Q7-Q8-Q9-Q 10 -Q 11 -Q 12 -Q 13 -Q 14 -Q 15 -Q 16 -Q 17 -Q 18 -Q 19 -Q 20 -Q 21 -Q 22 -Q 23 -Q 24 -Q 25 -Q 26 -Q 27 -Q 28 -Q 29 -Q 30 (Formula XVII) And Q1-Q 30 Their identities are provided in Table 18; Where XVIII is expressed as: R1-R2-R3-R4-R5-R6-R7-R8-R9-R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 (Formula XVIII) And R1-R 31 Their identities are provided in Table 19; Where XIX is represented as: S1-S2-S3-S4-S5-S6-S7-S8-S9-S 10 -S 11 -S 12 -S 13 -S 14 -S 15 -S 16 -S 17 -S 18 -S 19 -S 20 -S 21 -S 22 -S 23 -S 24 -S 25 -S 26 -S 27 -S 28 -S 29 -S 30 -S 31 -S 32 (Formula XIX) And S1-S 32 The identities are provided in Table 20; Where XX is represented as: T1-T2-T3-T4-T5-T6-T7-T8-T9-T 10 -T 11 -T 12 -T 13 -T 14 -T 15 -T 16 -T 17 -T 18 -T 19 -T 20 -T 21 -T 22 -T 23 -T 24 -T 25 -T 26 -T 27 -T 28 -T 29 -T 30 -T 31 -T 32 -T 33 (Formula XX) And T1-T 33 Their identities are provided in Table 21; Where XXI is represented as: U1]]U1-U2-U3-U4-U5-U6-U7-U8-U9-U 10 -U 11 -U 12 -U 13 -U 14 -U 15 -U 16 -U 17 -U 18 -U 19 -U 20 -U 21 -U 22 -U 23 -U 24 -U 25 -U 26 -U 27 -U 28 -U 29 -U 30 -U 31 -U 32 -U 33 -U 34 (Formula XXI) And U1-U 34 Their identities are provided in Table 22; Where equation XXII is represented as: V1-V2-V3-V4-V5-V6-V7-V8-V9-V 10 -V 11 -V 12 -V 13 -V 14 -V 15 -V 16 -V 17 -V 18 -V 19 -V 20 -V 21 -V 22 -V 23 -V 24 -V 25 -V 26 -V 27 -V 28 -V 29 -V 30 -V 31 -V 32 -V 33 -V 34 -V 35 (Formula XXII) And V1-V 35 Their identities are provided in Table 23; Where equation XXIII is represented as: W1-W2-W3-W4-W5-W6-W7-W8-W9-W 10 -W 11 -W 12 -W 13 -W 14 -W 15 -W 16 -W 17 -W 18 -W 19 -W 20 -W 21 -W 22 -W 23 -W 24 -W 25 -W 26 -W 27 -W 28 -W 29 -W 30 -W 31 -W 32 -W 33 -W 34 -W 35 -W 36 (Formula XXIII) And W1-W 36 Their identities are provided in Table 24; Where equation XXIV is represented as: X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 -X 17 -X 18 -X 19 -X 20 -X 21 -X 22 -X 23 -X 24 -X 25 -X 26 -X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 [[ID= And X1-X 37 Their identities are provided in Table 25; Where XXV is represented as: Y1 - Y2 - Y3 - Y4 - Y5 - Y6 - Y7 - Y8 - Y9 - Y 10 -Y 11 -Y 12 -Y 13 -Y 14 -Y 15 -Y 16 -Y 17 -Y 18 -Y 19 -Y 20 -Y 21 -Y 22 -Y 23 -Y 24 -Y 25 -Y 26 -Y 27 -Y 28 -Y 29 -Y 30 -Y 31 -Y 32 -Y 33 -Y 34 -Y 35 -Y 36 -Y 37 -Y 38 (Formula XXV) And Y1-Y 38 Their identities are provided in Table 26; Wherein, XXVI is represented as: Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 -Z 17 -Z 18 -Z 19 -Z 20 -Z 21 -Z 22 -Z 23 -Z 24 -Z 25 s -Z 26 -Z 27 -Z 28 -Z 29 -Z 30 -Z 31 -Z 32 -Z 33 -Z 34 -Z 35 -Z 36 -Z 37 -Z 38 -Z 39 (Formula XXVI) And Z1-Z 39 Their identities are provided in Table 27; Where equation XXVII is represented as: (AA)1-(AA)2-(AA)3-(AA)4-(AA)5-(AA)6-(AA)7-(AA)8-(AA)9-(AA) 10 -(AA) 11 -(AA) 12 -(AA) 13 -(AA) 14 -(AA) 15 -(AA) 16 -(AA) 17 -(AA) 18 -(AA) 19 -(AA) 20 -(AA) 21 -(AA) 22 -(AA) 23 -(AA) 24 -(AA) 25 -(AA) 26 -(AA) 27 -(AA) 28 -(AA) 29 -(AA) 30 -(AA) 31 -(AA) 32 -(AA) 33 -(AA) 34 -(AA) 35 -(AA) 36 -(AA) 37 -(AA) 38 -(AA) 39 -(AA) 40 (Formula XXVII) And (AA)1-(AA) 40 Their identities are provided in Table 28; Where equation XXVIII is represented as: (BB)1-(BB)2-(BB)3-(BB)4-(BB)5-(BB)6-(BB)7-(BB)8-(BB)9-(BB) 10 -(BB) 11 -(BB) 12 -(BB) 13 -(BB) 14 -(BB) 15 -(BB) 16 -(BB) 17 -(BB) 18 -(BB) 19 -(BB) 20 -(BB) 21 -(BB) 22 -(BB) 23 -(BB) 24 -(BB) 25 -(BB) 26 -(BB) 27 -(BB) 28 -(BB) 29 -(BB) 30 -(BB) 31 -(BB) 32 -(BB) 33 -(BB) 34 -(BB) 35 -(BB) 36 -(BB) 37 -(BB) 38 -(BB) 39 -(BB) 40 -(BB) 41 (Formula XXVIII) And (BB)1-(BB) 41 Their identities are provided in Table 29; Where XXIX is represented as: (CC)1-(CC)2-(CC)3-(CC)4-(CC)5-(CC)6-(CC)7-(CC)8-(CC)9-(CC) 10 -(CC) 11 -(CC) 12 -(CC) 13 -(CC) 14 -(CC) 15 -(CC) 16 -(CC) 17 -(CC) 18 -(CC) 19 -(CC) 20 -(CC) 21 -(CC) 22 -(CC) 23 -(CC) 24 -(CC) 25 -(CC) 26 -(CC) 27 -(CC) 28 -(CC) 29 -(CC) 30 -(CC) 31 -(CC) 32 -(CC) 33 -(CC) 34 -(CC) 35 -(CC) 36 -(CC) 37 -(CC) 38 -(CC) 39 -(CC) 40 -(CC) 41 -(CC) 42 (Formula XXIX) And (CC)1-(CC) 42 Their identities are provided in Table 30; Where the formula XXX is represented as: (DD)1-(DD)2-(DD)3-(DD)4-(DD)5-(DD)6-(DD)7-(DD)8-(DD)9-(DD) 10 -(DD) 11 -(DD) 12 -(DD) 13 -(DD) 14 -(DD) 15 -(DD) 16 -(DD) 17 -(DD) 18 -(DD) 19 -(DD) 20 -(DD) 21 -(DD) 22 -(DD) 23 -(DD) 24 -(DD) 25 -(DD) 26 -(DD) 27 -(DD) 28 -(DD) 29 -(DD) 30 -(DD) 31 -(DD) 32 -(DD) 33 -(DD) 34 -(DD) 35 -(DD) 36 -(DD) 37 -(DD) 38 -(DD) 39 -(DD) 40 -(DD) 41 -(DD) 42 -(DD) 43 (Formula XXX) And (DD)1-(DD) 43 Their identities are provided in Table 31; Wherein, XXXI is represented as: (EE)1-(EE)2-(EE)3-(EE)4-(EE)5-(EE)6-(EE)7-(EE)8-(EE)9-(EE) 10 -(EE) 11 -(EE) 12 -(EE) 13 -(EE) 14 -(EE) 15 -(EE) 16 -(EE) 17 -(EE) 18 -(EE) 19 -(EE) 20 -(EE) 21 -(EE) 22 -(EE) 23 -(EE) 24 -(EE) 25 -(EE) 26 -(EE) 27 -(EE) 28 -(EE) 29 -(EE) 30 -(EE) 31 -(EE) 32 -(EE) 33 -(EE) 34 -(EE) 35 -(EE) 36 -(EE) 37 -(EE) 38 -(EE) 39 -(EE) 40 -(EE) 41 -(EE) 42 -(EE) 43 -(EE) 44 (Formula XXXI) And (EE)1-(EE) 44 Their identities are provided in Table 32; Wherein, equation XXXII is represented as: (FF)1-(FF)2-(FF)3-(FF)4-(FF)5-(FF)6-(FF)7-(FF)8-(FF)9-(FF) 10 -(FF) 11 -(FF) 12 -(FF) 13 -(FF) 14 -(FF) 15 -(FF) 16 -(FF) 17 -(FF) 18 -(FF) 19 -(FF) 20 -(FF) 21 -(FF) 22 -(FF) 23 -(FF) 24 -(FF) 25 -(FF) 26 -(FF) 27 -(FF) 28 -(FF) 29 -(FF) 30 -(FF) 31 -(FF) 32 -(FF) 33 -(FF) 34 -(FF) 35 -(FF) 36 -(FF) 37 -(FF) 38 -(FF) 39 -(FF) 40 -(FF) 41 -(FF) 42 -(FF) 43 -(FF) 44 -(FF) 45 (Formula XXXII) And (FF)1-(FF) 45 Their identities are provided in Table 33; Where equation XXXIII is represented as: (GG)1-(GG)2-(GG)3-(GG)4-(GG)5-(GG)6-(GG)7-(GG)8-(GG)9-(GG) 10 -(GG) 11 -(GG) 12 -(GG) 13 -(GG) 14 -(GG) 15 -(GG) 16 -(GG) 17 -(GG) 18 -(GG) 19 -(GG) 20 -(GG) 21 -(GG) 22 -(GG) 23 -(GG) 24 -(GG) 25 -(GG) 26 -(GG) 27 -(GG) 28 -(GG) 29 -(GG) 30 -(GG) 31 -(GG) 32 -(GG) 33 -(GG) 34 -(GG) 35 -(GG) 36 -(GG) 37 -(GG) 38 -(GG) 39 -(GG) 40 -(GG) 41 -(GG) 42 -(GG) 43 -(GG) 44 -(GG) 45 -(GG) 46 (Formula XXXIII) And (GG)1-(GG) 46 Their identities are provided in Table 34; Wherein, XXXIV is represented as: (HH)1-(HH)2-(HH)3-(HH)4-(HH)5-(HH)6-(HH)7-(HH)8-(HH)9-(HH) 10 -(HH) 11 -(HH) 12 -(HH) 13 -(HH) 14 -(HH) 15 -(HH) 16 -(HH) 17 -(HH) 18 -(HH) 19 -(HH) 20 -(HH) 21 -(HH) 22 -(HH) 23 -(HH) 24 -(HH) 25 -(HH) 26 -(HH) 27 -(HH) 28 -(HH) 29 -(HH) 30 -(HH) 31 -(HH) 32 -(HH) 33 -(HH) 34 -(HH) 35 -(HH) 36 -(HH) 37 -(HH) 38 -(HH) 39 -(HH) 40 -(HH) 41 -(HH) 42 -(HH) 43 -(HH) 44 -(HH) 45 -(HH) 46 -(HH) 47 (Formula XXXIV) And (HH)1-(HH) 47 Their identities are provided in Table 35; Wherein, XXXV is represented as: (II)1-(II)2-(II)3-(II)4-(II)5-(II)6-(II)7-(II)8-(II)9-(II) 10 -(II) 11 -(II) 12 -(II) 13 -(II) 14 -(II) 15 -(II) 16 -(II) 17 -(II) 18 -(II) 19 -(II) 20 -(II) 21 -(II) 22 -(II) 23 -(II) 24 -(II) 25 -(II) 26 -(II) 27 -(II) 28 -(II) 29 -(II) 30 -(II) 31 -(II) 32 -(II) 33 -(II) 34 -(II) 35 -(II) 36 -(II) 37 -(II) 38 -(II) 39 -(II) 40 -(II) 41 -(II) 42 -(II) 43 -(II) 44 -(II) 45 -(II) 46 -(II) 47 -(II) 48 (Formula XXXV) And (II)1-(II) 48 The identities are provided in Table 36; Wherein, XXXVI is represented as: (JJ)1-(JJ)2-(JJ)3-(JJ)4-(JJ)5-(JJ)6-(JJ)7-(JJ)8-(JJ)9-(JJ) 10 -(JJ) 11 -(JJ) 12 -(JJ) 13 -(JJ) 14 -(JJ) 15 -(JJ) 16 -(JJ) 17 -(JJ) 18 -(JJ) 19 -(JJ) 20 -(JJ) 21 -(JJ) 22 -(JJ) 23 -(JJ) 24 -(JJ) 25 -(JJ) 26 -(JJ) 27 -(JJ) 28 -(JJ) 29 -(JJ) 30 -(JJ) 31 -(JJ) 32 -(JJ) 33 -(JJ) 34 -(JJ) 35 -(JJ) 36 -(JJ) 37 -(JJ) 38 -(JJ) 39 -(JJ) 40 -(JJ) 41 -(JJ) 42 -(JJ) 43 -(JJ) 44 -(JJ) 45 -(JJ) 46 -(JJ) 47 -(JJ) 48 -(JJ) 49 (Formula XXXVI) And (JJ)1-(JJ) 49 Their identities are provided in Table 37; Where equation XXXVII is represented as: (KK)1-(KK)2-(KK)3-(KK)4-(KK)5-(KK)6-(KK)7-(KK)8-(KK)9-(KK) 10 -(KK) 11 -(KK) 12 -(KK) 13 -(KK) 14 -(KK) 15 -(KK) 16 -(KK) 17 -(KK) 18 -(KK) 19 -(KK) 20 -(KK) 21 -(KK) 22 -(KK) 23 -(KK) 24 -(KK) 25 -(KK) 26 -(KK) 27 -(KK) 28 -(KK) 29 -(KK) 30 -(KK) 31 -(KK) 32 -(KK) 33 -(KK) 34 -(KK) 35 -(KK) 36 -(KK) 37 -(KK) 38 -(KK) 39 -(KK) 40 -(KK) 41 -(KK) 42 -(KK) 43 -(KK) 44 -(KK) 45 -(KK) 46 -(KK) 47 -(KK) 48 -(KK) 49 -(KK) 50 -(KK) 51 -(KK) 52 (Formula XXXVII) And (KK)1-(KK) 52 The identities are provided in Table 38; Wherein, equation XXXVIII is represented as: (LL)1-(LL)2-(LL)3-(LL)4-(LL)5-(LL)6-(LL)7-(LL)8-(LL)9-(LL) 10 -(LL) 11 -(LL) 12 -(LL) 13 -(LL) 14 -(LL) 15 -(LL) 16 -(LL) 17 -(LL) 18 -(LL) 19 -(LL) 20 -(LL) 21 -(LL) 22 -(LL) 23 -(LL) 24 -(LL) 25 -(LL) 26 -(LL) 27 -(LL) 28 -(LL) 29 -(LL) 30 -(LL) 31 -(LL) 32 -(LL) 33 -(LL) 34 -(LL) 35 -(LL) 36 -(LL) 37 -(LL) 38 -(LL) 39 -(LL) 40 -(LL) 41 -(LL) 42 -(LL) 43 -(LL) 44 -(LL) 45 -(LL) 46 -(LL) 47 -(LL) 48 -(LL) 49 -(LL) 50 -(LL) 51 -(LL) 52 -(LL) 53 -(LL) 54 -(LL) 55 (Formula XXXVIII) And (LL)1-(LL) 55 The identities are provided in Table 39; and Where the expression XXXIX is represented as: (MM)1-(MM)2-(MM)3-(MM)4-(MM)5-(MM)6-(MM)7-(MM)8-(MM)9-(MM) 10 -(MM) 11 -(MM) 12 -(MM) 13 -(MM) 14 -(MM) 15 -(MM) 16 -(MM) 17 -(MM) 18 -(MM) 19 -(MM) 20 -(MM) 21 -(MM) 22 -(MM) 23 -(MM) 24 -(MM) 25 -(MM) 26 -(MM) 27 -(MM) 28 -(MM) 29 -(MM) 30 -(MM) 31 -(MM) 32 -(MM) 33 -(MM) 34 -(MM) 35 -(MM) 36 -(MM) 37 -(MM) 38 -(MM) 39 -(MM) 40 -(MM) 41 -(MM) 42 -(MM) 43 -(MM) 44 -(MM) 45 -(MM) 46 -(MM) 47 -(MM) 48 -(MM) 49 -(MM) 50 -(MM) 51 -(MM) 52 -(MM) 53 -(MM) 54 -(MM) 55 -(MM) 56 -(MM) 57 -(MM) 58 -(MM) 59 (Formula XXXIX) And (MM)1-(MM) 59 Their identities are provided in Table 40.
7. The recombinant polypeptide of claim 5 or claim 6, wherein X1 comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 9, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39.
8. The recombinant polypeptide according to any one of claims 5 to 7, wherein the payload protein is a therapeutic peptide or protein.
9. A nucleic acid molecule encoding a signal peptide as described in any one of claims 1 to 4.
10. A nucleic acid molecule encoding a recombinant polypeptide as described in any one of claims 5 to 8.
11. A vector comprising the nucleic acid molecule as described in claim 9 or claim 10.
12. A cell comprising a nucleic acid molecule as claimed in claim 9 or claim 10, or a carrier as claimed in claim 11.
13. A composition comprising a nucleic acid molecule as claimed in claim 9 or claim 10, or a carrier as claimed in claim 11.
14. A method for treating a disease or condition in a subject in need, the method comprising administering to the subject an effective amount of the nucleic acid molecule as claimed in claim 10, thereby treating the disease or condition.
15. The method of claim 14, wherein the disease or condition is cancer.
16. A method for treating cancer in a subject in need, the method comprising administering to the subject a carrier comprising a nucleic acid molecule encoding a signal peptide fused to or linked with a payload protein, wherein the signal peptide is the signal peptide of any one of claims 1 to 4, and wherein the payload protein is a therapeutic peptide or protein that can be used to treat cancer.
Citation Information
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