Probing by chemoselective ligation uses bioorthogonal reactions to cross link two molecules without cross reactions with other biological functional groups. These chemical reactions permit regioselective covalent attachment of oligonucleotides with other biomolecules and permit the study of biomolecules real time in living systems without interfering normal cellular functions. Bioorthogonal probe "click" reactions are now widely used in chemical biology for many applications such as activity-based protein profiling, monitoring cell proliferation, generating novel enzyme inhibitors, and monitoring the synthesis of newly formed proteins. Other applications using this technique include protein targets, site-specific tagging of proteins, detection of DNA and RNA synthesis, detection of posttranslational modifications in proteins, and visualization of glycan non-invasive imaging in a living system.

Bio-Synthesis offers oligo synthesis and modifications for bioconjugation based on bioorthogonal chemistry. These oligo bioorthogonal probes can be modified at 5’, 3’, or internally during chemical synthesis or conjugate post-synthetically.We have provided hundreds of DNA or RNA conjugates using a variety of bioorthogonal chemical reactions such as photoinduced cycloaddition of diaryl tetrazole to alkene, 1, 3-dipolar cycloaddition between azides and cyclooctynes, also known as copper-free click chemistry, strain promoted alkyne-azide cycloaddition (SPAAC), oxime/hydrazone formation between aldehyde and ketones etc. Contact Bio-Synthesis for oligonucleotide bioorthogonal probes.

Bio-Synthesis offers a number of bioorthogonal reactions that can be use for detection, tracking and imaging purposes. Our services include but not limited to: click chemistry, Staudinger ligation, hydrazide-aldehyde reactions, aminoxy-aldehyde reactions and others.

Contact us to start your Bioorthogonal Probes

Diel-Alder Reaction

Diel-alder reaction consists of a 4 + 2 cycloaddition between a diene and an alkene, often called dienophile. Diene modified oligonucleotides can be prepared by solid-phase synthesis using a 3,5-hexadiene phosphoramidite derivative, which could be incorporated into the oligo at the 5' end.

Diel-Alder Reaction
Modification 5' Int 3' Purification
Diene-modified oligonucleotides Y Dual HPLC

Tetrazine TCO Ligation Chemistry

The Trans-Cyclooctene-Tetrazine click chemistry is a very powerful tool in catalyst‐free bioconjugation chemistry. The reaction follows an inverse‐electron demand Diels‐Alder cycloaddition reaction of trans‐cyclooctenes (TCO) with tetrazines. This bioorthogonal reaction possesses an exceptionally high reaction speed and nearly quantitative yield under physiocological conditions with reaction rate constants 2000 M-1 s−1 (in 9:1 methanol/water). The extremely fast kinetics and selectivity enables the conjugation of two low abundance biopolymers in an aqueous and otherwise complex chemical environment through the formation of a stable dihydropyridazine. This bioorthogonal reaction possesses extreme selectivity and biocompatibility, such that the complimentary reagents can form covalent bonds within richly functionalized biological systems, in some cases, living organisms.

Tetrazine TCO Ligation Chemistry
Modification 5' Int 3' Purification
TCO-PEG12 NHS Y -- -- Single HPLC
Tetrazine-PEG4 NHS Y Y Y Dual HPLC
Tetrazine-PEG5 NHS Y Y Y Dual HPLC

Hydrazine Aldehyde Reaction

The reaction between an aldehyde or ketone and a hydrazide or hydrazine functional group to form a hydrzone bond has been frequently used for bioconjugation reaction. Aldehyde, hydrazide modified oligonucleotides can be prepared by solid-phase synthesis or functionalized post-synthetically.

Modification 5' Int 3' Purification
Aldehyde Y HPLC
Hydrazide Y Y Y Dual HPLC

Aminooxy Aldehyde Reaction

The chemoselective conjugation of an aminooxy modified oligo with an aldehyde is similar to that of the reaction of a hydrazide with an aldehyde, except instead of giving a hydrazone bond it yields an oxime linkage. Aminooxy modified oligonucleotides can be prepared by solid-phase synthesis using a 5'-aminooxy modifier 11. This modifier is based on a tetraethylene glycol linkage for improved solubility and for reducing the potential negative impact on hybridization of the oligo. The oxime formed from the reaction of alkyloxyamines with aldehydes creates a stable covalent bond.

Modification 5' Int 3' Purification
5' aminooxy 11 modification Y HPLC

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