CN1108820C - 单表位重复或多联单表位重复-表位疫苗的制备方法 - Google Patents
单表位重复或多联单表位重复-表位疫苗的制备方法 Download PDFInfo
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- 229960005486 vaccine Drugs 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims description 20
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 37
- 229920001184 polypeptide Polymers 0.000 claims abstract description 30
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 30
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 12
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 10
- 239000002671 adjuvant Substances 0.000 claims abstract description 9
- 102000014914 Carrier Proteins Human genes 0.000 claims abstract description 3
- 108010078791 Carrier Proteins Proteins 0.000 claims abstract description 3
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- 238000010168 coupling process Methods 0.000 claims description 12
- 238000005859 coupling reaction Methods 0.000 claims description 12
- 241000555268 Dendroides Species 0.000 claims description 6
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 6
- 230000036039 immunity Effects 0.000 claims description 2
- 244000052769 pathogen Species 0.000 abstract description 11
- 230000001717 pathogenic effect Effects 0.000 abstract description 8
- 239000000126 substance Substances 0.000 abstract description 3
- 230000028993 immune response Effects 0.000 abstract 1
- 230000006798 recombination Effects 0.000 abstract 1
- 238000005215 recombination Methods 0.000 abstract 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 5
- 230000004044 response Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 3
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- 239000004411 aluminium Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000009849 deactivation Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 229940031626 subunit vaccine Drugs 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229940028617 conventional vaccine Drugs 0.000 description 1
- KZNICNPSHKQLFF-UHFFFAOYSA-N dihydromaleimide Natural products O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229960002317 succinimide Drugs 0.000 description 1
- -1 succinimide ester Chemical class 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
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Abstract
本发明属生物工程技术领域,本发明首先人工合成带有病原体天然蛋白或基因重组蛋白上的特定表位的表位多肽;其表位是以重复的形式出现在人工合成的表位多肽上;其表位多肽耦联到载体蛋白或载体多肽上形成耦联物或者直接合成为树枝状多肽分子;其耦联物或树枝状多肽分子配以适当佐剂制备成单表位重复或多联单表位重复-表位疫苗。本发明可诱导表位特异性的免疫应答;可对付因病原体变异而导致的疫苗失效;可制备出多联疫苗。
Description
本发明属于生物工程技术领域,特别涉及表位疫苗的制备和生产。
常规疫苗制备技术通常采用以下方法:
(1)利用减毒或灭活的病原体(如脊髓灰质炎病毒);
(2)利用病原体的天然的或基因重组的相关组分,如细菌类毒素、病毒包膜蛋白等制备的亚单位疫苗;
(3)利用病毒的核酸制备疫苗。
现有的这些疫苗制备技术在预防由变异频度很高的某些病原体(如爱滋病病毒和流感病毒)引发的传染病时,存在着很大的困难;此外,现有的疫苗不能直接诱导预先确定的表位特异性的免疫应答。
本发明的目的是为克服已有技术的不足之处,设计出的一种单表位重复-表位疫苗的制备方法,使其具有不需要减毒或灭活的病原体或病原体的天然蛋白或基因重组蛋白抗原以及病原体的核酸;能够直接诱导预先确定的表位特异性的免疫应答;能够对付因病原体变异而导致的疫苗失效;能够制备出预防不同病原体的多联疫苗等突出的优势。
本发明提出的单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
(1)人工合成带有HIV(病原体)天然蛋白或基因重组蛋白上的特定表位的表位多肽;
(2)所说的表位是以重复的形式出现在人工合成的表位多肽上;
(3)所说的表位多肽耦联到载体蛋白或载体多肽上形成耦联物或者直接合成为树枝状多肽分子(MAP,Multiple Antigen Peptide);
(4)所说的耦联物或树枝状多肽分子配以适合的佐剂制备成单表位重复的表位疫苗或者是制备成多联单表位重复-表位疫苗。
本发明提出的另一种单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成带有HIV天然蛋白或基因重组蛋白上的特定表位的表位多肽;
2)所说的表位是以重复的形式出现在人工合成的表位多肽上;
3)所说的表位多肽直接合成为树枝状多肽分子;
4)所说的树枝状多肽分子配以适合的佐剂制备成单表位重复的表位疫苗或者是制备成多联单表位重复-表位疫苗。
本发明提出的又一种单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成带有HIV天然蛋白或基因重组蛋白上的几种特定表位的表位多肽;
2)将所说的几种单表位重复的表位多肽混合,利用MBS将几种表位多肽与载体蛋白BSA同时耦联;
3)将耦联物分别和佐剂混合制备成多联单表位重复的表位疫苗。
本发明根据由本发明人最先提出的表位疫苗的全新理论(Immunology Today,20:588-589,1999年12月)设计出的表位疫苗的制备方法具有如下效果:
第一、能诱导很强的针对特定表位的免疫应答,与相应的亚单位疫苗相比,能显著提高针对特定表位的抗体水平(10-20倍);
第二、某些天然蛋白上的特定表位在机体内不能诱导出较强的具有防护作用的免疫应答,利用本发明制备的疫苗能够诱导出高滴度的表位特异性的抗体(表位特异性的抗体量为10-240微克每毫升血清),能够起到有效的免疫防护作用;
第三、应用本发明制备的多联单表位-表位疫苗能够在机体内同时诱导出针对多个特定表位的抗体应答。
实施例一:制备HIV-1单表位重复的表位疫苗
步骤1.人工合成以下三种重复表位的表位多肽:
C(ELDKWA)G(ELDKWA)G(ELDKWA)G(ELDKWA)
C(GPGRAFY)(GPGRAFY)(GPGRAFY)(GPGRAFY)
C(RILVERYLKD)GG(RILVERYLKD)
步骤2.利用MBS(mmaleimidobenzoyl-N-hydroxy succinimide ester)分别将三种表位多肽与载体蛋白BSA耦联;
步骤3.将三种耦联物分别和铝佐剂混合制备成三种单表位重复的表位疫苗。
实施例二制备HIV-1多联单表位重复的表位疫苗
方法1:步骤1、2、3与实施例一相同;
步骤4.将上述制备的三种单表位重复的表位疫苗混合免疫;
方法2:步骤1与实施例一相同;
步骤2将上述制备的三种单表位重复的表位多肽混合,利用MBS(mmaleimidobenzoyl-N-Arydroxy succinimide ester)将三种表位多肽与载体蛋白BSA同时耦联;
步骤3将耦联物分别和铝佐剂混合制备成三种单表位重复的表位疫苗。
Claims (4)
1.一种单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成带有HIV天然蛋白或基因重组蛋白上的特定表位的表位多肽;
2)所说的表位是以重复的形式出现在人工合成的表位多肽上;
3)所说的表位多肽耦联到载体蛋白或载体多肽上形成耦联物;
4)所说的耦联物配以适合的佐剂制备成单表位重复的表位疫苗或者是制备成多联单表位重复-表位疫苗。
2.一种单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成带有HIV天然蛋白或基因重组蛋白上的特定表位的表位多肽;
2)所说的表位是以重复的形式出现在人工合成的表位多肽上;
3)所说的表位多肽直接合成为树枝状多肽分子;
4)所说的树枝状多肽分子配以适合的佐剂制备成单表位重复的表位疫苗或者是制备成多联单表位重复-表位疫苗。
3.如权利要求1或2所述的表位疫苗的制备方法,其特征在于还包括:
将所说的制备的几种单表位重复的表位疫苗混合免疫制备成多联单表位重复-表位疫苗。
4.一种单表位重复-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成带有HIV天然蛋白或基因重组蛋白上的几种特定表位的表位多肽;
2)将所说的几种单表位重复的表位多肽混合,利用MBS将几种表位多肽与载体蛋白BSA同时耦联;
3)将耦联物分别和佐剂混合制备成多联单表位重复的表位疫苗。
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CN99123807A CN1108820C (zh) | 1999-11-12 | 1999-11-12 | 单表位重复或多联单表位重复-表位疫苗的制备方法 |
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CA2558733C (en) * | 2004-02-06 | 2016-04-19 | Inserm (Institut National De La Sante Et De La Recherche Medicale) | A polypeptide derived from gp41, a vaccine composition comprising said polypeptide, and uses for treating an infection by an hiv virus in an individual |
CN104710537A (zh) * | 2015-04-09 | 2015-06-17 | 汪运山 | 柯萨奇病毒ca16 vp1重组抗原及其单克隆抗体与应用 |
CN106892974B (zh) * | 2017-03-07 | 2020-09-15 | 中国医科大学 | 一种基于肿瘤抗原ecm1的长肽ere1及其在肿瘤免疫治疗中的应用 |
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WO1986007383A1 (en) * | 1985-06-04 | 1986-12-18 | Biotechnology Research Partners, Ltd. | Autoantigen vaccines |
WO1994003208A1 (en) * | 1992-07-30 | 1994-02-17 | Yeda Research And Development Company Ltd. | Conjugates of poorly immunogenic antigens and synthetic peptide carriers and vaccines comprising them |
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WO1986007383A1 (en) * | 1985-06-04 | 1986-12-18 | Biotechnology Research Partners, Ltd. | Autoantigen vaccines |
WO1994003208A1 (en) * | 1992-07-30 | 1994-02-17 | Yeda Research And Development Company Ltd. | Conjugates of poorly immunogenic antigens and synthetic peptide carriers and vaccines comprising them |
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