Development and Uses of TFP-dPEG®4-biotinidase-resistant biotin
For an explanation of biotinylation with dPEG® products, please click here.
Biotinidase is an amidohydrolase that breaks the covalent bond between biotin and amino acids such as lysine (i.e., biocytin). In vivo, biotinidase recycles biotin. Although necessary for healthy bodily function, biotinidase potentially creates problems when biotinylated proteins are used in serological assays, since endogenous biotinidase removes the biotin from these proteins.
Dr. D. Scott Wilbur and colleagues at the University of Washington in Seattle developed biotinidase-resistant biotin in 2006. Development of biotinidase-resistant biotin arose from a need for biotinylation reagents that could facilitate in vivo antibody pretargeting. However, to improve the product’s water solubility, Quanta BioDesign conjugated a short amino-dPEG®4-propionic acid linker to biotinidase-resistant biotin. For practical use in bioconjugation, the TFP ester then was synthesized.
Like our other dPEG® biotinylation products, TFP-dPEG®4-biotinidase-resistant biotin will not cause aggregation and precipitation of biomolecules to which it is conjugated. Moreover, the dPEG® spacer in this product reduces non-specific binding. Aggregation, precipitation, and non-specific binding are common problems with the widely used LC-biotin reagents.
TFP-dPEG®4-biotinidase-resistant biotin resists amide hydrolysis because the non-natural amino acid to which it is conjugated prevents the active site of biotinidase from hydrolyzing the amide bond connecting biotin to the dPEG®4 spacer. This product potentially is beneficial for applications that use serological fluids containing endogenous biotinidase. Useful roles for TFP-dPEG®4-biotinidase-resistant biotin include the following:
- therapeutic applications such as antibody pretargeting with biotin-streptavidin systems;
- supramolecular constructs (g., nanoparticles) used in vivo that depend on biotin-(strept)avidin affinity to maintain structural integrity; and,
- ELISAs of serological fluids and tissue samples using biotin-(strept)avidin affinity.
Products Similar to TFP-dPEG®4-biotinidase-resistant biotin
In addition to this TFP ester-activated construct, Quanta BioDesign offers a more extended version of this product and an equivalent-length product functionalized as the N-hydroxysuccinimidyl (NHS) ester. TFP esters possess greater hydrolytic stability and amine reactivity compared to NHS esters.
Commercial Scale Production Is Available for TFP-dPEG®4-biotinidase-resistant biotin
If you need bulk product in a larger package size than our standard sizes, please contact us for a quote. Our commercial capabilities permit us to manufacture this product at any scale that you need.
Hydrophobic LC-biotin reagents are known to be problematic in bioconjugation work. Moreover, when working with serological fluids and tissue samples, endogenous biotinidase potentially creates problems when analyzing results.
Stop using inferior biotinylation reagents! You can do better. Quanta BioDesign offers better biotinylation tools. Our dPEG®-based biotinylation reagents offer water solubility, improved hydrodynamic volume, no background noise (which means better signal), and no protein precipitation caused by aggregation. Why would you not use something better?
For better biotinylation, click the “Add to Cart” button now. You will not regret it.