Description

TFP-dPEG®12-biotinidase-resistant biotin, product number QBD-10204, is a water-soluble biotinylation reagent modified to resist hydrolysis by biotinidase (EC 3.5.1.12). This product combines Biotinoyl-2-aminobutyric acid with the 2,3,5,6-tetrafluorophenyl (TFP) ester of a medium-length, single molecular weight, discrete polyethylene glycol (dPEG®) linker. TFP-dPEG®12-biotinidase-resistant biotin retains biotin’s typical high affinity for avidin and streptavidin but also blocks the action of biotinidase on biotin. This product is useful in applications such as ELISAs of tissue samples and antibody pretargeting, where endogenous biotinidase can recycle biotin.

Biotinidase is an amidohydrolase that recycles biotin in vivo by breaking the covalent bond between biotin and amino acids such as lysine (i.e., biocytin). 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 to facilitate in vivo antibody pretargeting. To improve the product’s water solubility and ability to reach the binding pocket of avidin, a medium-length amino-dPEG®12-propionic acid linker was conjugated to the biotinidase-resistant biotin and subsequently functionalized as an TFP ester.

Like our other dPEG® biotinylation products, TFP-dPEG®12-biotinidase-resistant biotin, QBD-10204, will not cause aggregation and precipitation of biomolecules to which it is conjugated, and the dPEG® spacer reduces non-specific binding. It is potentially beneficial for applications using serological fluids that contain endogenous biotinidase. Useful roles for TFP-dPEG®12-biotinidase-resistant biotin include the following:
therapeutic applications such as antibody pretargeting with biotin-streptavidin systems;
supramolecular constructs (e.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.

Specifications

Documents

References

1. Greg T. Hermanson, Bioconjugate Techniques, 3rd Edition, Elsevier, Waltham, MA 02451, 2013, ISBN 978-0-12-382239-0; See Chapter 18, Discrete PEG Reagents, pp. 787-821, for a full overview of the dPEG® products.

Description

TFP-dPEG®4-biotinidase-resistant biotin, product number QBD-10203, is a water-soluble biotinylation reagent modified to resist hydrolysis by biotinidase (EC 3.5.1.12). This product combines Biotinoyl-2-aminobutyric acid with the 2,3,5,6-tetrafluorophenyl (TFP) ester of a single molecular weight, discrete polyethylene glycol (dPEG®) linker. TFP-dPEG®4-biotinidase-resistant biotin retains biotin’s typical high affinity for avidin and streptavidin but also blocks the action of biotinidase on biotin. This product is useful in applications such as ELISAs of tissue samples and antibody pretargeting, where endogenous biotinidase can recycle biotin.
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 to facilitate in vivo antibody pretargeting. However, to improve the product’s water solubility, a short amino-dPEG®4-propionic acid linker was conjugated to the biotinidase-resistant biotin and subsequently functionalized as an TFP ester. This product is essentially the same as QBD-10202, but the TFP ester is more stable to hydrolysis in aqueous media and has a higher optimal pH range of 7.5 – 8.5 compared to NHS esters.

Like our other dPEG® biotinylation products, TFP-dPEG®4-biotinidase-resistant biotin, QBD-10203, will not cause aggregation and precipitation of biomolecules to which it is conjugated, and the dPEG® spacer reduces non-specific binding.

TFP-dPEG®4-biotinidase-resistant biotin resists amide hydrolysis because of the non-natural amino acid to which it is conjugated. It is potentially beneficial for applications using serological fluids that contain 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 (e.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.

Specifications

Documents

References

1. Greg T. Hermanson, Bioconjugate Techniques, 3rd Edition, Elsevier, Waltham, MA 02451, 2013, ISBN 978-0-12-382239-0; See Chapter 18, Discrete PEG Reagents, pp. 787-821, for a full overview of the dPEG® products.

Description

NHS-dPEG®4-biotinidase-resistant biotin, product number QBD-10202, is a water-soluble biotinylation reagent modified to resist hydrolysis by biotinidase (EC 3.5.1.12). This product combines Biotinoyl-2-aminobutyric acid with the N-hydroxysuccinimidyl (NHS) ester of a single molecular weight, discrete polyethylene glycol (dPEG®) linker. NHS-dPEG®4-biotinidase-resistant biotin retains biotin’s typical high affinity for avidin and streptavidin. However, the modified biotin prevents recycling by biotinidase. This product is useful in applications such as ELISAs of tissue samples and antibody pretargeting, where endogenous biotinidase can recycle biotin.
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 to facilitate in vivo antibody pretargeting. However, to improve the product’s water solubility, a short amino-dPEG®4-propionic acid linker was conjugated to the biotinidase-resistant biotin and subsequently functionalized as an NHS ester.

Like our other dPEG® biotinylation products, NHS-dPEG®4-biotinidase-resistant biotin will not cause aggregation and precipitation of biomolecules to which it is conjugated, and the dPEG® spacer reduces non-specific binding.

NHS-dPEG®4-biotinidase-resistant biotin resists amide hydrolysis because of the non-natural amino acid to which it is conjugated. It is potentially beneficial for applications using serological fluids that contain endogenous biotinidase. Useful roles for NHS-dPEG®4-biotinidase-resistant biotin include the following:
therapeutic applications such as antibody pretargeting with biotin-streptavidin systems;
supramolecular constructs (e.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.

Specifications

Documents

References

1. Greg T. Hermanson, Bioconjugate Techniques, 3rd Edition, Elsevier, Waltham, MA 02451, 2013, ISBN 978-0-12-382239-0; See Chapter 18, Discrete PEG Reagents, pp. 787-821, for a full overview of the dPEG® products.
2. Biotin Reagents for Antibody Pretargeting. 7. Investigation of Chemically Inert Biotinidase Blocking Functionalities for Synthetic Utility. D. Scott Wilbur, Donald K. Hamlin, and Ming-Kuan Chyan. Bioconjugate Chem. 2006, 17 (6) pp 1514–1522. October 28, 2006. DOI: 10.1021/bc060084m.