StreptaClick® Precision
for Assay Development
StreptaClick® Precision — Monovalently Binding Streptavidin for Click Chemistry
Customize StreptaClick® to match your precise assay or product requirements. Our novel StreptaClick® Precision molecule, specifically designed for Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) or copper-free Strain-promoted azide-alkyne cycloaddition (SPAAC) click chemistry has exactly one azide or DBCO reactive site. This enables the precise attachment of exactly one molecule of choice to each StreptaClick® Precision molecule, catering to applications requiring a stoichiometrically precise 1:1 ratio of reporter to binder. Furthermore, StreptaClick® Precision maintains its remarkable biotin-binding capability and effortlessly connects to any biotinylated antibody or protein in solution without causing aggregation via its two clustered biotin binding sites in cis-position. The dual binding sites ensure biotin binding is not blocked by bulky molecules such as oligonucleotides.
Possible applications include:
- DNA origami, precise labeling of reaction surfaces or beads
- Super-resolution microscopy
- Antibody-oligonucleotide conjugate based immunoassays such as immuno-PCR, immuno-RCA, and Proximity Ligation Assays
- Prototyping or screening antibody-drug conjugates, label free antibody-antigen interaction kinetics for antibody therapeutics
- Screening targeting antibodies conjugated to delivery systems such as lipid nanoparticles or extracelluar vesicles in the development of advanced therapies and medicinal products (ATMPs).
- Antigen-labeling to measure receptor/ligand or antigen/antibody interactions.
Should you require a greater ratio of reporters per binder we are currently developing StreptaClick® Precision versions with exactly three or seven reactive groups. If you wish to explore novel applications using StreptaClick®, please do not hesitate to contact us.
StreptaClick® Precision - a cis-divalent streptavidin primed for Click Chemistry and multiplexing
StreptaClick® Precision differs from our previous monovalent StreptaClick® Color and StreptaClick® HRP molecules in that it has been redesigned with dual biotin binding sites clustered in cis-position, making it cis-divalent. Nonetheless, this design still exhibits monovalent binding properties to antibodies while conserving its native binding affinity to biotin, ensuring unmatched specificity in molecular interactions. In early experiments we observed that monovalent StreptaClick® could have its biotin binding obscured by bulky molecules such as oligonucleotides, a problem that has now been solved with StreptaClick® Precision. Furthermore, the cis position of the two biotin binding sites still negates antibody aggregation and allows mixing in solution just as our other monovalent StreptaClick® products. This makes StreptaClick® Precision ideal for multiplexing.
How it works
Easily attach the azide labeled StreptaClick® Precision molecule to your reporter molecule of choice via click chemistry. The choice of azide or DBCO functionalization enables either Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) or copper-free Strain-promoted azide-alkyne cycloaddition (SPAAC) click chemistry, ensuring accurate and specific labeling in many different types of experiments.
StreptaClick® Precision Workflow
Achieve high conjugation efficiency
Conjugating reporters to StreptaClick® Precision via CuAAC or SPAAC click chemistry typically results in 90-100% labeling efficiency with only 1.3X excess of reporter to StreptaClick® molecules.
Native gel comparing StreptaClick® conjugated to oligonucleotides to controls
- StreptaClick® Precision – Oligo (off-the shelf kit, 14 bp)
- Control: StreptaClick® Precision – DBCO
- StreptaClick® Precision – DBCO + 43 bp oligo – azide
- StreptaClick® Precision – DBCO + 41 bp oligo – azide
- StreptaClick® Precision – DBCO + 37 bp oligo – azide
- Control: StreptaClick® Precision – Azide
- StreptaClick® Precision – Azide + 20 bp oligo – DBCO
- StreptaClick® Precision – Azide + 20 bp oligo – DBCO
Attach StreptaClick® Precision conjugate to any biotinylated antibody or surface
After the click chemistry reaction, add the newly formed StreptaClick® Precision construct to your biotinylated protein or surface of interest for near immediate conjugation. We also have a convenient biotinylation kit available for purchase, BiotinPureTM, if you lack access to biotinylated antibodies.
This innovation grants you full control over labeling position and ratio, enabling detailed exploration of protein behaviors, interactions, and localization.
The image below shows an example of a fresh frozen mouse spleen tissue in which StreptaClick® Precision – 1 Oligo bound to an anti B220 biotinylated antibody from Biolegend interacts with B220 proteins on B-lymphocytes. The interaction is visualized via immunofluorescence imaging by hybridization of an antisense oligo with a 647nm fluorochrome to StreptaClick® Precision – 1 Oligo. Other possible readouts for antibody-oligonucleotide conjugate based methods include q-PCR and DNA sequencing.
The perfect starting point for assay development
StreptaClick Precision has greatly facilitated our research based on lipid nanoparticles (LNP).
We study subsets of macrophages using antibody coated LNPs containing functional RNA to
genetically reprogram the subsets through antibody-targeted delivery. We can now simply
generate a large batch of StreptaClick Precision LNPs via click chemistry and then easily mix
this with different biotinylated antibodies from our library to screen for the most suitable
antibodies to use for the targeting.
We have recently evaluated a new version of StreptaClick Precision that features, in
addition to azide for click chemistry, a tracer fluorochrome. This allows us to confirm LNP
uptake by the correct subset as well as correlate this to the expression of target RNA by GFP
or mCherry in another fluorescent channel.
StreptaClick Precision has become a game changer for us as it is convenient, consistent
and flexible compared to directly conjugating each antibody to the LNPs, and also cost
effective.
Anders Etzerodt, Associate Professor, PhD
Department of Biomedicine
Aarhus University, Denmark