寡核苷酸Oligo和Antibody抗體偶聯(lián)技術(shù)MagicLink

中文原理介紹：

寡核苷酸可以通過硫醇-馬來酰亞胺和腙偶聯(lián)方法與抗體/蛋白質(zhì)偶聯(lián)。硫醇-馬來酰亞胺方法通常使用雙功能接頭（例如 SMCC）進(jìn)行。然而，SMCC修飾的蛋白質(zhì)極不穩(wěn)定，并且經(jīng)常自反應(yīng)，因?yàn)樗ǔ：邪泛土虼蓟鶊F(tuán)，導(dǎo)致大量的同源交聯(lián)。當(dāng)存在其他硫醇基團(tuán)時(shí)，共軛鍵可交換。在腙偶聯(lián)方法中，共軛反應(yīng)基于肼和醛之間的反應(yīng)。盡管使用了催化劑，但這種方法的收率很低。分子的大小和官能團(tuán)的不可接近性導(dǎo)致產(chǎn)量低。此外，聯(lián)動(dòng)穩(wěn)定性也是一個(gè)問題。

BroadPharm（國內(nèi)總代理：靶點(diǎn)科技）的MagicLink™技術(shù)通過結(jié)合PEG和專有的交聯(lián)化學(xué)解決了所有問題。MagicLink™ 化學(xué)試劑盒和試劑盒根本不需要 TCEP、DTT 或催化劑！

首先，抗體與LINK NHS酯反應(yīng)，將LINK引入抗體中。此外，胺修飾的寡核苷酸與MAGIC NHS酯反應(yīng)，將MAGIC引入寡核苷酸結(jié)構(gòu)中。其次，在純化兩個(gè)反應(yīng)產(chǎn)物后，它們立即相互反應(yīng)，產(chǎn)生具有穩(wěn)定環(huán)連接的抗體-寡核苷酸偶聯(lián)物。

- 新穎性：第三代蛋白質(zhì)交聯(lián)技術(shù)

- 產(chǎn)率：偶聯(lián)效率在95-100%左右

- 速率：即時(shí)反應(yīng);整個(gè)過程不到1小時(shí)

- 鍵穩(wěn)定性：市場上穩(wěn)定的共價(jià)鍵偶聯(lián)物

- 一致性：批次間重現(xiàn)性好

- 靈活：以所需的比例使用您自己的寡核苷酸和抗體

- 保質(zhì)期：在室溫下長時(shí)間、穩(wěn)定的凍干船

- 抗體選擇：任何蛋白質(zhì)/抗體、抗體片段

- 寡核苷酸選擇：10~120個(gè)堿基的單鏈寡核苷酸，最多80個(gè)堿基對(duì)的雙鏈寡核苷酸; 連接化學(xué)在 5 端和 3 端都有效

- 無需偶聯(lián)后凈化步驟

- 包括陽性對(duì)照抗體和寡核苷酸，可確認(rèn)方案成功

該試劑盒還可用于將寡核苷酸與堿性磷酸酶（AP）、辣根過氧化物酶（HRP）、牛血清白蛋白（BSA）、大豆過氧化物酶（SBP）、青霉素酶（PNC）、鏈霉親和素及其偶聯(lián)物、α-D-半乳糖苷酶、葡萄糖-t-磷酸脫氫酶等偶聯(lián)。

這種寡核苷酸-酶偶聯(lián)物（“寡酶"）可用于多種雜交和檢測(cè)形式，包括斑點(diǎn)印跡、南方/北方印跡、原位和溶液雜交/捕獲方案。

原理示意圖：

英文原理介紹：

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.

應(yīng)用舉例：

訂購信息：

名稱：MagicLink™ Oligo Antibody Conjugation Kit

貨號(hào)：BP-50004
規(guī)格：3*100ug

總代理：靶點(diǎn)科技