Compositional and structural heterogeneity of avidin glycopeptides

Bruch, Richard C.; White, Harold B.

doi:10.1021/bi00264a033

Cited by 101 publications

(67 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another commonly used protein in BioFET studies is Avidin, 30,36,71 which is related to streptavidin, has an affinity for biotin but has a higher pI and is therefore oppositely charged at the physiological pH. This protein however is a poor choice for comparative study for several reasons, firstly, electrophoresis reveals distinctive charge heterogeneity even in purified avidin and both compositional and structural heterogeneity due to its carbohydrate content, 84 and secondly, its higher pI can result in higher non-specific binding due to a larger net-charge in physiological solutions. 85 Another biomolecule commonly used in BioFET experiments is prostate-specific antigen (PSA) due to biomedical applications as a prostate cancer biomarker, however the biomolecular capture step is more complex than streptavidin, requiring the use of a complex biomolecule such as an antibody or aptamer.…”

Section: Streptavidin Biochemistrymentioning

confidence: 99%

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Lowe

Sun

Zeimpekis

et al. 2017

Analyst

131

107

View full text Add to dashboard Cite

a Field-Effect Transistor sensors (FET-sensors) have been receiving increasing attention for biomolecular sensing over the last two decades due to their potential for ultra-high sensitivity sensing, label-free operation, cost reduction and miniaturisation. Whilst the commercial application of FET-sensors in pH sensing has been realised, their commercial application in biomolecular sensing (termed BioFETs) is hindered by poor understanding of how to optimise device design for highly reproducible operation and high sensitivity. In part, these problems stem from the highly interdisciplinary nature of the problems encountered in this field, in which knowledge of biomolecular-binding kinetics, surface chemistry, electrical double layer physics and electrical engineering is required. In this work, a quantitative analysis and critical review has been performed comparing literature FET-sensor data for pH-sensing with data for sensing of biomolecular streptavidin binding to surface-bound biotin systems. The aim is to provide the first systematic, quantitative comparison of BioFET results for a single biomolecular analyte, specifically streptavidin, which is the most commonly used model protein in biosensing experiments, and often used as an initial proofof-concept for new biosensor designs. This novel quantitative and comparative analysis of the surface potential behaviour of a range of devices demonstrated a strong contrast between the trends observed in pH-sensing and those in biomolecule-sensing. Potential explanations are discussed in detail and surfacechemistry optimisation is shown to be a vital component in sensitivity-enhancement. Factors which can influence the response, yet which have not always been fully appreciated, are explored and practical suggestions are provided on how to improve experimental design.

show abstract

Section: Streptavidin Biochemistrymentioning

confidence: 99%

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Lowe

Sun

Zeimpekis

et al. 2017

Analyst

131

107

View full text Add to dashboard Cite

show abstract

“…Avidin contains twice the number of the basic amino acids lysine and arginine than SA; therefore, the isoelectric point of AV is approximately 10 compared with about 7.0 for SA. Approximately 10% of avidin's mass is due to heterogeneous oligosaccharides, largely composed of mannose and N-acetylglucosamine (DeLange, 1970;Bruch and White, 1982). Because of their high affinity for biotin, they have been used in numerous biotechnological applications Bayer, 1988, 1989) and in different in vivo procedures such as radioimmunodetection and drug immunotargeting (Hnatowich et al, 1987;Paganelli et al, 1990Paganelli et al, , 1995.…”

mentioning

confidence: 99%

Biochemical modifications of avidin improve pharmacokinetics and biodistribution, and reduce immunogenicity

Chinol

Casalini²,

Maggiolo³

et al. 1998

Br J Cancer

110

View full text Add to dashboard Cite

Summary Pretargeting techniques using the avidin-biotin system have shown encouraging results in both diagnostic and therapeutic clinical trials. It has been shown that in cancer therapy the ideal agent to be used for pretargeting should have a plasma half-life longer than avidin and lower immunogenicity than streptavidin in order for these procedures to be applied safely and repeatedly in patients. We prepared a recombinant form of avidin with no carbohydrates and avidins, biochemically modified either by decreasing the positive charges with succinic anhydride or by linking polyethylene glycol (PEG) at three different molar ratios and evaluated their in vivo behaviour after i.p. administration in mice. The succinylation and PEGylation of avidin increased the plasma half-life proportionally to the degree of protein modification. The procedures, however, affected the biotin binding to some extent. The biodistribution studies showed that, for all six time points (ranging from 20 min to 18 h post-injection), the liver and kidney to blood ratios were lower for PEGylated avidins than native, recombinant and succinyl avidin. Recombinant and low PEGylated avidin evoked an immune response in all mice after at least three injections. Native, recombinant and succinyl avidins showed higher serum titres than PEGylated avidins. In conclusion, the conjugation of avidin to PEG chains (n = 7) originates a compound with a suitable blood clearance, low immunogenicity and concurrent low cross-reactivity with avidin.

show abstract

“…This suggested that the binding capacity of the complex was maintained during its transport through the epithelial monolayer. In fact, avidin has a resistance to proteolytic digestion due to the presence of 10 % carbohydrates [22].…”

Section: Discussionmentioning

confidence: 99%

Paracellular Transport of Avidin Saturated or not with Biotinylated Cobalamin through Caco-2 Cell Epithelium Monolayer

Meyer

Pons

Namour

et al. 2001

Cell Physiol Biochem

View full text Add to dashboard Cite

Background: The cationic charge of molecules may promote their uptake across epithelia, which are rich in brush border anionic sites. The transport of unsaturated avidin and avidin saturated with a biotinylated compound was investigated across Caco-2 adenocarcinoma cell with fetal enterocyte phenotype. Methods: The unsaturated avidin and avidin saturated with either biotin or a biotinyl-cobalamin conjugate (biotinyl-Cbl) were iodinated to follow their transport through the cell monolayer. Their apparent permeability coefficient (Papp) and transepithelial pathway were determined and compared to those for control radiolabeled markers [³H]-mannitol, [¹²⁵I]-β-lactoglobulin and [⁵⁷Co]-cobalamin/intrinsic factor (Cbl/IF). Results: The Papp of [¹²⁵I]-avidin estimated at 2.8 x 10^-7 ± 0.08 cm/ s was close to that for mannitol that uses paracellular pathway. The binding of biotin or biotin conjugate to avidin enhanced its tetrameric conformation. The Papp for [¹²⁵I]-avidin/biotin and [¹²⁵I]- avidin/biotinyl-Cbl were respectively increased by 2-fold, compared to that for [¹²⁵I]-avidin and 4-fold, compared to that for [¹²⁵I]-β-lactoglobulin and [⁵⁴Co]-Cbl/IF. The protein was not accumulated in the cell and was found in intact form in the basolateral side, after its transport across the monolayer. Chloroquine (0.66 µmol/ml) did not significantly decrease the Papp for [¹²⁵I]-avidin/biotinyl-Cbl. Conversely it decreased by 80% the Papp for Cbl/IF, that uses transepithelial pathway. Conclusions: Avidin (either saturated or not with biotin and biotinyl-Cbl) was able to cross the monolayer of Caco-2 cell line through a paracellular pathway. This study pointed out the interest for using this protein as a shuttle for increasing the transport rate of biotinylated compounds through fetal epithelial barriers.

show abstract

Compositional and structural heterogeneity of avidin glycopeptides

Cited by 101 publications

References 46 publications

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Biochemical modifications of avidin improve pharmacokinetics and biodistribution, and reduce immunogenicity

Paracellular Transport of Avidin Saturated or not with Biotinylated Cobalamin through Caco-2 Cell Epithelium Monolayer

Contact Info

Product

Resources

About