MagicLink™ Oligo Antibody Conjugation Kit

Product Citations

1. Wang, T., Wang, X., Luo, S., Zhang, P., Li, N., Chen, C., ... & Huang, R. P. (2024). Constructions, Purifications and Applications of DNA-Antibody Conjugates: A Review. ACS omega, 9(49), 47951-47963.
   https://pubs.acs.org/doi/full/10.1021/acsomega.4c07714

• Overview

  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.

• Components

  1. MAGIC NHS (MW ~900)
  2. LINK NHS (MW ~900)
  3. Oligo control,
  4. Antibody control
  5. Protein concentrator
  6. Reaction buffer

• Application

  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 arrays

  Oligozymes application: dot blots, Southern/Northern blots, In situ, and solution hybridization/capture

• Images

  

  

• Protocol

  MagicLink™ Oligonucleotide Antibody Conjugation Kit

• Reference

  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