背景介绍
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.

工作原理

产品组分

1. MAGIC NHS (MW ~900)
2. LINK NHS (MW ~900)
3. oligo control,
4. antibody control
5. protein concentrator
6.reaction buffer
产品应用
MagicLink Oligo antibody Labeling Kits are used to conjugate oligo with primary antibody, secondary antibodies, enzymes, etc. for the bioassays below. Oligo-antibody application:
*Immuno-Polymerase Chain Reaction (immuno-PCR)
*Proximity Ligation Assay (PLA)
*Electrochemical Proximity Assay (ECPA)
*Multiplexed Assay Development
*Proximity extension assay
*3DNA technology in lateral flow
*3DNA technology in cellular targeting
*single cell CITE-seq experiments
*DNA-PAINT imaging
protein arraysOligozymes application: dot blots, Southern/Northern blots, In situ, and solution hybridization/capture

参考文献
1.McMahon NP, Jones JA, Kwon S, et al. Oligonucleotide conjugated antibodies permit highly multiplexed immunofluorescence for future use in clinical histopathology. J Biomed Opt. 2020;25(5):1-18. doi:10.1117/1.JBO.25.5.056004
2.Wiener, J., Kokotek, D., Rosowski, S. et al. Preparation of single- and double-oligonucleotide antibody conjugates and their application for protein analytics. Sci Rep 10, 1457 (2020). https://doi.org/10.1038/s41598-020-58238-6

点击化学寡肽抗体偶联试剂盒MagicLink™ Oligo Antibody Conjugation Kit

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，并且经常具有自反应性，因为它通常含有胺和硫醇基团，会导致大量的同型交联。当存在其他硫醇基团时，共轭键是可交换的。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，但该方法收率低。分子的大小和官能团的不可接近性导致低产率。此外，连锁稳定性是一个问题。

BroadPharm 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™ 化学品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。此外，胺修饰的寡核苷酸与 MAGIC NHS 酯反应以将 MAGIC 引入寡核苷酸结构中。其次，在两个反应性伙伴纯化后，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：共轭效率在 95-100% 左右

*速率：即时反应；整个过程不到1小时

*连接稳定性：市场上稳定必选的共价键结合物

*一致性：批次间重现性好

*灵活：以您想要的比例使用您自己的寡核苷酸和抗体

*保质期：环境温度下长、稳定的冷冻干燥船

*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，以及长达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

*无需进行偶联后清理步骤

*包含阳性对照抗体和寡核苷酸以确认方案成功

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)、霉素酶 (PNC)、链霉亲和素及其偶联物、α-D-半乳糖苷酶偶联、葡萄糖-t-磷酸脱氢酶等。

此类寡核苷酸-酶缀合物(“寡酶”)可用于多种杂交和检测形式，包括斑点印迹、Southern/Northern印迹、原位和溶液杂交/捕获方案。