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InvivoGen的CpG ODNs 优势

· 涵盖最全的CpG寡聚脱氧核苷酸 (CpG ODN)或细菌DNA

· 生物学活性检测

· 随产品提供去内毒素水

· 高溶解度

CpG ODNs 是合成的单链DNA短片段,含有特定的非甲基化CpG二核苷酸序列。CpG ODNs含有部分或全部磷硫酰骨架(phosphorothioated (PS) backbone)。与细菌基因组DNA的磷酸二酯骨架相对(phosphodiester (PO) backbone)。

三个主要刺激性CpG ODNs的分类,是基于其结构特性与在人外周血单核细胞(PBMC),某些B细胞和 类浆树突细胞(pDC)的活性而划分。三类CpG ODNs是Class A (Type D), Class B (Type K) 和Class C。


Classes of CpG ODNs

- CpG-A ODNs 以含有CpG回文序列为中西的PO骨架和PS修饰的3’端poly-G链构成。CpG-A ODNs刺激类浆树突细胞(pDC)可以较强的产生IFN-a,但是会较弱的激活TLR9依赖的NF-kB信号通路,产生较少的促炎性细胞因子(如白细胞介素6)。

- CpG-B ODNs 是包含一个或多个CpG二核苷酸的完整PS骨架结构序列。CpG-B ODNs可以较强的活化B细胞和TLR9介导的NF-kB信号,但刺激分泌IFN-a的效果较弱。

- CpG-C ODNs 结构是Class A和B的混合体。它包含了完整的PS骨架和CpG回文序列。CpG-C ODNs可以较强的引起pDC分泌IFN-a,在刺激B细胞时也可以得到较好的效果。

Class A

Class A (type D) CpG ODNs (CpG-A ODNs) 以含有CpG回文序列为中西的PO骨架和PS修饰的3’端poly-G链构成。

Poly G 末端在分子间形成四分体,从而产生高分子量结构。这样的结构增强了稳定性与内涵体的胞内转运,促使大量IFN-a的产生和pDCs的成熟。

CpG-A ODNs也是NK细胞的强活化剂,其作用是通过间接的细胞因子信号来达到活化效果。

Ballas ZK. et al., 1996. Induction of NK activity in murine and human cells by CpG motifs in oligodeoxynucleotides and bacterial DNA. J Immunol. 157(5):1840-5. Krug A. et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN-alpha/beta in plasmacytoid dendritic cells. Eur J Immunol. 31(7):2154-63. Verthelyi D. et al., 2001. Human peripheral blood cells differentially recognize and respond to two distinct CPG  motifs. J Immunol. 166(4):2372-7.

 

Class A CpG ODNs 刺激 HEK-Blue™ hTLR9 细胞 (货号:hkb-htlr9)

用不同浓度的A-class CpG ODNs, ODN 1585, ODN 2216 或 ODN 2236及相应的对照ODNs 刺激HEK-Blue™ hTLR9 细胞 (表达人源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

 

 

Class A CpG ODNs 刺激 HEK-Blue™ mTLR9 细胞 (货号:mkb-htlr9)

用不同浓度的A-class CpG ODNs, ODN 1585, ODN 2216 或 ODN 2236及相应的对照ODNs 刺激HEK-Blue™ mTLR9 细胞 (表达鼠源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

Class B

Class B (type K) CpG ODNs (CpG-B ODNs)是18-28摩尔的线性脱氧寡核苷酸。CpG-B ODNs 是包含一个或多个6摩尔CpG二核苷酸的完整PS骨架结构序列。人源的最佳序列是GTCGTT[1],鼠源的最佳序列是GACGTT[2]。

CpG-B ODNs是B细胞和NK细胞的强刺激剂,对于IFN-a分泌的刺激效果却较弱[3,4]。CpG-B ODNs表现出较好的抗肿瘤效果以及Th1型佐剂的活性。

 Hartmann G & Krieg AM., 2000. Mechanism and function of a newly identified CpG DNA motif in human primary B cells. J Immunol. 164(2):944-53. 2- Krieg AM. et al., 1995. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature. 374(6522):546-9. 3- Hartmann G. et al., 2000. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primate immune responses in vitro and in vivo. J Immunol. 164(3):1617-24. 4- Gürsel M. et al., 2002. Differential and competitive activation of human immune cells by distinct classes of CpG oligodeoxynucleotide. J Leukoc Biol. 71(5):813-20.

 


Class B CpG ODNs 刺激 HEK-Blue™ hTLR9 细胞 (货号:hkb-htlr9)

用不同浓度的B-class CpG ODNs, ODN 1668, ODN 1826, ODN 2006 或 ODN 2007及相应的对照ODNs 刺激HEK-Blue™ hTLR9 细胞 (表达人源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

 

 

Class B CpG ODNs 刺激 HEK-Blue™ mTLR9 细胞 (货号:mkb-htlr9)

用不同浓度的B-class CpG ODNs, ODN 1668, ODN 1826, ODN 2006 或 ODN 2007及相应的对照ODNs 刺激HEK-Blue™ mTLR9 细胞 (表达鼠源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

Class C

Class C CpG ODNs (CpG-C ODNs)结构是Class A和B的混合体。它包含了完整的PS骨架和CpG回文序列。

C-Class CpG ODNs可以较强的引起pDC分泌IFN-a,在刺激B细胞时也可以得到较好的效果。C-Class CpG ODNs是很强的Th1-诱导性佐剂。

Vollmer J. et al., 2004. Characterization of three CpG oligodeoxynucleotide classes with distinct immunostimulatory activities. Eur J Immunol. 34(1):251-62.



Class C CpG ODNs 刺激 HEK-Blue™ hTLR9 细胞 (货号:hkb-htlr9)

用不同浓度的C-class CpG ODNs, ODN 2395 或 ODN M362及相应的对照ODNs 刺激HEK-Blue™ hTLR9 细胞 (表达人源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

 


Class C CpG ODNs 刺激 HEK-Blue™ mTLR9 细胞 (货号:mkb-htlr9)

用不同浓度的C-class CpG ODNs, ODN 2395 或 ODN M362及相应的对照ODNs 刺激HEK-Blue™ mTLR9 细胞 (表达鼠源TLR9和NF-κB诱导 SEAP 报告基因, 刺激物与HEK-Blue Detection试剂共孵育24 小时, NF-ΚB-诱导 SEAP的水平在OD 655 nm检测。

 

正确选择ODN类型

Class A CpG ODNs Class B CpG ODNs Class C CpG ODNs
应用: 应用: 应用:
刺激 pDCs 刺激 B 细胞 混合CpG A和CpG B的功能
诱导 IFN-a 产生 活化NF-kB
疫苗佐剂
产品: 产品: 产品:
ODN 2216 (human preferred) ODN 1826 (mouse) ODN 2395 (human/mouse)
ODN 1585 (mouse) ODN 1668 (mouse) ODN M362 (human/mouse)
ODN 2336 (human preferred) ODN 2006 (human preferred)
ODN BW006 (human/mouse)
ODN 2007 (Bovine/porcine)
ODN D-SL01 (Multispecies)

Class Backbone B cells pDCs IFN-α Il-6
A PS-PO + ++++ ++++ +
B PS ++++ ++++ + +++
C PS +++ +++ +++ ++

 

ODN的品质经专业的实验室检测,并发表文献



实验结果表明,从InvivoGen购买的CpG-C, 不同批次的产品刺激细胞并产生IgG的效果均一稳定,而从Sigma 和Coley购买的产品活性则非常不连续


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  136. /www/new_annoron/vendor/topthink/think-template/src/template/driver/File.php ( 2.41 KB )
  137. /www/new_annoron/vendor/topthink/think-template/src/template/contract/DriverInterface.php ( 0.86 KB )
  138. /www/new_annoron/runtime/temp/392bd12e4c3a40f421a0bf2a7ca1ae7d.php ( 13.12 KB )
  139. /www/new_annoron/vendor/topthink/framework/src/think/Response.php ( 8.81 KB )
  140. /www/new_annoron/vendor/topthink/framework/src/think/response/Html.php ( 0.97 KB )
  141. /www/new_annoron/vendor/topthink/framework/src/think/Cookie.php ( 6.06 KB )
  142. /www/new_annoron/vendor/topthink/think-trace/src/Html.php ( 4.42 KB )
  1. CONNECT:[ UseTime:0.000422s ] mysql:host=127.0.0.1;port=3306;dbname=www_annoron_com;charset=utf8
  2. SHOW FULL COLUMNS FROM `ecs_article` [ RunTime:0.000919s ]
  3. SELECT * FROM `ecs_article` WHERE `article_id` = 651 LIMIT 1 [ RunTime:0.000406s ]
  4. SHOW FULL COLUMNS FROM `ecs_article_cat` [ RunTime:0.000809s ]
  5. SELECT * FROM `ecs_article_cat` WHERE `cat_id` = 5 LIMIT 1 [ RunTime:0.000322s ]
  6. SELECT * FROM `ecs_article_cat` WHERE `parent_id` = 11 [ RunTime:0.000306s ]
0.027639s