For more information, contact our technical representative for more questions on SoluLink products at 888.625.0670 or email us at info@solulink.com.
SoluLinK’s HyNic conjugation linker family of products have the advantages of being super stable in solution, they have a built-in verification and UV-traceability, is controllable and it is not susceptible to non-specific binding, making it superior to the standard older methods of conjugation. Specifically, the method readily reacts with primary amines on a protein (ε-amino group of lysine) via an NHS-ester.
The built-in traceable HyNic group (hydrazinonicotinate) in the conjugated proteins or other biomolecules that provides a confirmation and quantification of the conjugation. HyNic-modified proteins then react to form stable conjugates in the presence of other (aromatic) aldehyde-modified proteins or biomolecules. For more information check out the Solulink catalog or request information about SoluLinK’s conjugation services solulink@solulink.com.
SoluLinK references and citations are listed here.
Additional advantages:
-Stable- forms a covalent hydrazone bond (> 94oC 2hr)
-Traceable- spectral confirmation of conjugate formation and purification
-Conjugation Control- molar substitution ratio (MSR) readily determined
-Biocompatabile- reducing agents not required (e.g. cyanoborohydride)
-Specificity- reacts only with aromatic aldehydes in the presence of -NH2, -SH,-COOH and other protein functionalities
Previous Methods
For the last 30 years, there have been only three methods commonly used to prepare biomacromolecule conjugates.
1. Amide formation using an amine and an activated carboxylic acid.
2. Thiol ether or disulfide formation using a thiol and either a maleimide or a
pyridiyldisulfide, respectively.
3. Biotin/avidin coupling using the ligand biotin and an avidin to its receptor
protein.
Each of these has significant disadvantages that have limited the development of many types of products and diagnostic assays. Solulink offers a novel technology that does not suffer from limited stability of incorporated reactive functionalities (i.e. maleimide and thiol chemistries) or non-specific binding (i.e. biotin/avidin), yet retains the positive aspects, such as conjugation efficiency.
Solulink’s chemistry is simple in concept. It is based on the reaction in water of 6-hydrazino- nicotinamide (HyNic) moieties with carbonyl moieties to yield a stable hydrazone bond. The chemistry shown here can be engineered to link small molecules (e.g. peptides, fluorophores), biomolecules (e.g. proteins, oligonucleotides, DNA), or solid surfaces (e.g. glass, plastic, latex, or silica beads).
Unlike other small molecule conjugation methods, molecules or surfaces modified with either HyNic or carbonyl moieties have extended stabilities in aqueous environments. These moieties can be incorporated on any surface and remain reactive without special handling requirements. The hydrazone formed by these partners is stable, in contrast to the hydrazones formed by more commonly accessible hydrazides that form unstable acyl hydrazones. Acyl hydrazones require reduction to stabilize the bond.
The Solulink Bioconjugation System’s unique ability to couple molecules together without the addition of a coupling reagent can be readily applied to the following systems:
-protein/protein conjugation
-protein/oligonucleotide conjugation
-oligonucleotide/oligonucleotide conjugation
-cDNA/fluorophore conjugation
-oligonucleotide/bead conjugation
-oligonucleotide/fluorophore conjugation
-5’-hydrazine or 5’-aldehyde primer extension and conjugation
Unlike maleimido/thiol chemistry that requires immediate use, biomolecules or surfaces modified with Solulink’s hydrazine/carbonyl reactive moieties can be prepared, stored for longer periods, and reacted when required.
Uses:
1. Small molecule chemistry that is relatively inexpensive.
2. Biomolecules and surfaces modified with Solulink’s reactive moieties,
hydrazines, and carbonyls, have extended stabilities (>3 months).
3. Modification incorporation can be readily quantified colorimetrically.
4. The reactive moieties do not interact with functionalities on biomolecules.
5. The chemistry is all done in aqueous buffered media, yielding high efficiency
conjugation.
6. Applicable to all current diagnostic assays, including gene and protein micro-
arrays. This technology will be a “plug-in” and not require significant
development time to become a part of assays available today. New
protocols/technologies not possible with current methods will be possible.
7. Solulink’s reactive moieties can be incorporated during solid phase synthesis of
The products offered here are for research use only. Any commercial application will require a license from Solulink. The Solulink Conjugation System is patented and has additional patents pending. Please contact Solulink for information regarding licensing information.
No license is granted or implied to any patents to technologies for which the end user applies our products.
