CN1108819C - 抗变异-表位疫苗的制备方法 - Google Patents
抗变异-表位疫苗的制备方法 Download PDFInfo
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- CN1108819C CN1108819C CN 99123806 CN99123806A CN1108819C CN 1108819 C CN1108819 C CN 1108819C CN 99123806 CN99123806 CN 99123806 CN 99123806 A CN99123806 A CN 99123806A CN 1108819 C CN1108819 C CN 1108819C
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- 229960005486 vaccine Drugs 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims description 12
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 25
- 229920001184 polypeptide Polymers 0.000 claims abstract description 24
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 24
- 239000002671 adjuvant Substances 0.000 claims abstract description 6
- 108091007433 antigens Proteins 0.000 claims abstract description 6
- 102000036639 antigens Human genes 0.000 claims abstract description 5
- 102000014914 Carrier Proteins Human genes 0.000 claims abstract description 3
- 108010078791 Carrier Proteins Proteins 0.000 claims abstract description 3
- 230000008878 coupling Effects 0.000 claims description 5
- 238000010168 coupling process Methods 0.000 claims description 5
- 238000005859 coupling reaction Methods 0.000 claims description 5
- 239000000412 dendrimer Substances 0.000 claims description 5
- 229920000736 dendritic polymer Polymers 0.000 claims description 5
- 230000035772 mutation Effects 0.000 claims description 5
- 244000052769 pathogen Species 0.000 abstract description 10
- 230000001717 pathogenic effect Effects 0.000 abstract description 9
- 239000000126 substance Substances 0.000 abstract description 2
- 230000028993 immune response Effects 0.000 abstract 1
- 230000004044 response Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 3
- 241000555268 Dendroides Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 241000713772 Human immunodeficiency virus 1 Species 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
- 238000000034 method 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
- 102000004169 proteins and genes Human genes 0.000 description 2
- 229940031626 subunit vaccine Drugs 0.000 description 2
- LLXVXPPXELIDGQ-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-(2,5-dioxopyrrol-1-yl)benzoate Chemical compound C=1C=CC(N2C(C=CC2=O)=O)=CC=1C(=O)ON1C(=O)CCC1=O LLXVXPPXELIDGQ-UHFFFAOYSA-N 0.000 description 1
- 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
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940028617 conventional vaccine Drugs 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
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- 238000003786 synthesis reaction Methods 0.000 description 1
- 241000712461 unidentified influenza virus Species 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一个或一个以上抗原蛋白上的一个或一个以上特定关键表位上的几个限制性突变型表位的表位多肽;突变型表位以一个或重复的形式出现在人工合成的一条多肽上;表位多肽耦联到载体蛋白或载体多肽或者利用MAP技术***直接合成为树枝状分子上;表位多肽的耦联物配以适当佐剂制备成抗变异—表位疫苗。本发明能诱导表位特异性的免疫应答;能对付因病原体变异而导致的疫苗失效;能制备多联疫苗。
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)利用病原体的核酸制备疫苗。
现有的这些疫苗制备技术在预防由变异频率很高的某些病原体(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。
本发明的目的是为了解决或部分解决病原体变异导致疫苗失效的世界性难题,设计出的一种抗变异-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。
本发明提出的抗变异-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成一个或一个以上抗原蛋白上的一个或一个以上特定关键表位上的几个限制性突变型表位的表位多肽;
2)所说的一个或一个以上突变型表位是以一个或以重复的形式出现在人工合成的一条多肽(即多突变型表位-表位多肽)上;
3)这个表位多肽耦联到载体蛋白或载体多肽或者利用MAP(Multiple AntigenPeptide)技术***直接合成为树枝状分子上;
4)表位多肽的耦联物或树枝状分子配以适当佐剂制备成抗变异-表位疫苗或抗变异-多表位-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,应用本发明制备的抗变异-多表位-表位疫苗能够在机体内同时诱导出针对多个限制性突变型变异表位的抗体应答。
实施例一:制备HIV-1抗变异-多表位-表位疫苗
步骤1.合成分别含有四个和三个限制性突变型的两种抗变异-表位多肽
CG(ELDKWA)G(ELEKWA)G(ELNKWA)G(ELDNWA)
CG(GPGRAFY)G(GPGKAFY)G(GPGQAFY)
步骤2.利用MBS(m-maleimidobenzoyl-N-hydroxy succinimide ester)将两种抗变异表位多肽
混和后与载体蛋白BSA耦联,或者与BSA分别耦联后混合;
步骤3.将耦联物分别和佐剂混合制备成抗变异-表位疫苗。
实施例二:采用树枝状表位多肽分子制备HIV-1抗变异-表位疫苗
步骤1.采用直接合成法利用MAP(Multiple Antigen Peptide)技术***直接合成
含一个特定表位ELDKWA的四种限定性突变型的树枝状分子上;
步骤2.将合成的树枝状表位多肽分子与适当佐剂混合制备成抗变异-表位疫
苗。
Claims (2)
1、一种抗变异-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成一个或一个以上抗原蛋白上的一个或一个以上特定关键表位上的几个限制性突变型表位的表位多肽;
2)所说的或多个突变型表位是以一个或以重复的形式出现在人工合成的一条多肽上;
3)所说的表位多肽耦联到载体蛋白或载体多肽;
4)所说的表位多肽的耦联物配以适当佐剂制备成抗变异-表位疫苗或抗变异-多表位-表位疫苗。
2、一种抗变异-表位疫苗的制备方法,其特征在于包括以下步骤:
1)人工合成一个或一个以上抗原蛋白上的一个或一个以上特定关键表位上的几个限制性突变型表位的表位多肽;
2)所说的一个或一个以上突变型表位是以一个或以重复的形式出现在人工合成的一条多肽上;
3)所说的表位多肽利用MAP技术***直接合成为树枝状分子上;
4)所说的表位多肽的树枝状分子配以适当佐剂制备成抗变异-表位疫苗或抗变异-多表位-表位疫苗。
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CN 99123806 CN1108819C (zh) | 1999-11-12 | 1999-11-12 | 抗变异-表位疫苗的制备方法 |
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CN 99123806 CN1108819C (zh) | 1999-11-12 | 1999-11-12 | 抗变异-表位疫苗的制备方法 |
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CN1256149A CN1256149A (zh) | 2000-06-14 |
CN1108819C true CN1108819C (zh) | 2003-05-21 |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9196800B2 (en) | 1996-06-26 | 2015-11-24 | Osram Gmbh | Light-radiating semiconductor component with a luminescence conversion element |
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CN102391375A (zh) * | 2011-10-24 | 2012-03-28 | 武汉伊艾博科技有限公司 | 蛋白抗原多个多肽抗原表位偶联载体生产抗体的方法 |
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- 1999-11-12 CN CN 99123806 patent/CN1108819C/zh not_active Expired - Fee Related
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9196800B2 (en) | 1996-06-26 | 2015-11-24 | Osram Gmbh | Light-radiating semiconductor component with a luminescence conversion element |
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