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寡核苷酸Oligo和Antibody抗体偶联技术MagicLink

发布时间:2022-12-04   点击次数:314次

中文原理介绍:

寡核苷酸可以通过硫醇-马来酰亚胺和腙偶联方法与抗体/蛋白质偶联。硫醇-马来酰亚胺方法通常使用双功能接头(例如 SMCC)进行。然而,SMCC修饰的蛋白质极不稳定,并且经常自反应,因为它通常含有胺和硫醇基团,导致大量的同源交联。当存在其他硫醇基团时,共轭键可交换。在腙偶联方法中,共轭反应基于肼和醛之间的反应。尽管使用了催化剂,但这种方法的收率很低。分子的大小和官能团的不可接近性导致产量低。此外,联动稳定性也是一个问题。

BroadPharm(国内总代理:靶点科技)的MagicLink™技术通过结合PEG和专有的交联化学解决了所有问题。MagicLink™ 化学试剂盒和试剂盒根本不需要 TCEP、DTT 或催化剂!

首先,抗体与LINK NHS酯反应,将LINK引入抗体中。此外,胺修饰的寡核苷酸与MAGIC NHS酯反应,将MAGIC引入寡核苷酸结构中。其次,在纯化两个反应产物后,它们立即相互反应,产生具有稳定环连接的抗体-寡核苷酸偶联物。

- 新颖性:第三代蛋白质交联技术

- 产率:偶联效率在95-100%左右

- 速率:即时反应;整个过程不到1小时

- 键稳定性:市场上稳定的共价键偶联物

- 一致性:批次间重现性好

- 灵活:以所需的比例使用您自己的寡核苷酸和抗体

- 保质期:在室温下长时间、稳定的冻干船

- 抗体选择:任何蛋白质/抗体、抗体片段

- 寡核苷酸选择:10~120个碱基的单链寡核苷酸最多80个碱基对的双链寡核苷酸; 连接化学在 5 端和 3 端都有效

- 无需偶联后净化步骤

- 包括阳性对照抗体和寡核苷酸,可确认方案成功

该试剂盒还可用于将寡核苷酸与碱性磷酸酶(AP)、辣根过氧化物酶(HRP)、牛血清白蛋白(BSA)、大豆过氧化物酶(SBP)、青霉素酶(PNC)、链霉亲和素及其偶联物、α-D-半乳糖苷酶、葡萄糖-t-磷酸脱氢酶等偶联。

这种寡核苷酸-酶偶联物(“寡酶")可用于多种杂交和检测形式,包括斑点印迹、南方/北方印迹、原位和溶液杂交/捕获方案。


原理示意图:




英文原理介绍:

Oligonucleotide can be conjugated to antibody/protein by thiol-maleimide and hydrazone coupling approaches. The thiol-maleimide approach is commonly performed with a bifunctional linker (such as SMCC). However, SMCC-modified protein is extremely unstable and often self-reactive since it often contains both amine and thiol groups that cause a significant amount of homo-crosslinking. The conjugation linkage is exchangeable when other thiol groups are present. In the hydrazone coupling approach, the conjugation reaction is based on a reaction between hydrazine and aldehyde. This method is low yielding although a catalyst is used. The size of the molecule and inaccessibility of the functional group contribute to low yield. In addition, linkage stability is an issue.

BroadPharm's MagicLink™ technology solves all the problems by combining PEG and proprietary crosslinking chemistry. MagicLink™ chemistry and kit do not need TCEP, DTT, or catalysts at all!
First, antibody reacts with LINK NHS ester to introduce LINK into the antibody. Also, amine modified oligonucleotide reacts with MAGIC NHS ester to introduce MAGIC into the oligonucleotide structure. Second, after purification of both reactive partners, they instantly react with each other yielding an antibody-oligonucleotide conjugate with a stable ring linkage.

- Novelty: 3rd generation protein crosslink technology

Yield: conjugation efficiency around 95-100%

Rate: instant reaction; the whole process less than 1 hour

Linkage stability: most stable covalent bonded conjugates on the market

Consistence: good reproducibility from batch to batch

Flexible: use your own oligo and antibody at your desired ratio

Shelf life: long, stable freeze dried ship at ambient temperature

Antibody selection: any proteins/antibody, antibody fragment

Oligo selection: single strand oligos of 10 120 bases, and double strand oligos up to 80 base pairs;

  Linking chemistry works at both 5 and 3 end

Post-conjugation clean-up step not necessary

Positive control antibody and oligo included to enable confirmation of protocol success

The kit can also be used to conjugate oligonucleotides to alkaline phosphatase (AP), horseradish peroxidase (HRP), bovine serum albumin (BSA), soybean peroxidase (SBP), penicillinase (PNC), streptavidin and its conjugates, alpha-D-galactosidase, glucose-t-phosphate dehydrogenase, etc.
Such oligonucleotide-enzyme conjugates ("oligozymes") can be used in a variety of hybridization and detection formats, including dot blots, Southern/Northern blots, in situ and solution hybridization/capture schemes.


应用举例:

订购信息:

名称:MagicLink™ Oligo Antibody Conjugation Kit

货号:BP-50004
规格:2*100ug

总代理:靶点科技

靶点科技(北京)有限公司

靶点科技(北京)有限公司

地址:中关村生命科学园北清创意园2-4楼2层

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