Glycosylation of Bovine Serum Albumin with D-[14C]-Glucose

Kublashvili, R. I.; Ugrekhelidze, D.

doi:10.1007/s10600-005-0144-5

Cited by 3 publications

(4 citation statements)

References 5 publications

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“…To evaluate the potential of aluminum oxide nanoparticles as a carrier for immunization, we first derivatized Al 2 O 3 by silanization to provide them with a terminal amine group. The amination percentage was 2%, which is similar to the theoretical proportion of primary amines for BSA (1.2%) 30 and higher than the theoretical proportion for ovalbumin (OVA) (<0.7%). On the other hand, the amine load was 32 μmol/g aluminum oxide, which is comparable to that of silanized Al 2 O 3 particles (16 μmol/g) that have been previously described.…”

Section: ■ Results and Discussionsupporting

confidence: 65%

Aluminum Oxide Nanoparticles as Carriers and Adjuvants for Eliciting Antibodies from Non-immunogenic Haptens

et al. 2012

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Functionalized nanomaterials have applications yet to be discovered, especially in the biological field and, in particular, the in vivo production of antibodies. In this paper, we show that aluminum oxide nanoparticles that are covalently coupled to haptens (small molecular mass compounds) activate the immune system, eliciting antibodies (polyclonal and monoclonal) specific for the herbicide atrazine, the antibiotic sulfasalazine, and the vitamin biotin in mice and rabbits. The particles play the role of carrier and adjuvant, with the immune response being dependent on size and crystallinity. The affinity constants of the antibodies are similar to those reached with traditional immunization strategies based on the use of carrier proteins. This approach has not been previously described, being of scientific and practical interest, as it can lead to immunogens safer than the conventional ones, potentially applicable to human vaccination. In addition, the useful advantages of this technique include the stability of the metallic particles, the synthesis of immunogens in organic media, the versatility of particle derivatization, the ease of purification, the full chemical characterization of immunogens, the lack of a requirement for an adjuvant, the reduction of the cross reactivity, and the low cost of materials. Also, the analytical performances of the immunoassays developed using these antibodies are comparable to those obtained by the standard protocols. Analytical applications of the developed ELISAs were fully demonstrated. Characterization (IgG nature, affinity constants, etc.) of the immunoreagents were also fully assessed.

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Section: ■ Results and Discussionsupporting

confidence: 65%

Aluminum Oxide Nanoparticles as Carriers and Adjuvants for Eliciting Antibodies from Non-immunogenic Haptens

et al. 2012

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“…Often, the applied temperature determines which path of the reaction route prevails, complicating the comparison of results obtained at different temperatures . High temperatures, especially when applied to a system at high pH, have been found to be the most important factor enhancing the Maillard reaction. ,, The kinetics shown in Figure also indicate that the rate of BSA glucosylation decreases over time, causing a “tailing” of the kinetic curve. Martins and Van Boekel attributed this to the decrease in pH over time, due to the formation of acidic products, which consequently slows the Maillard reaction.…”

Section: Resultsmentioning

confidence: 99%

“…The BSA−Glc mixtures were prepared by dissolving 2.5 mg/mL BSA and 25 mg/mL Glc in 0.05 M bicine buffer (#039K5437, Sigma) adjusted to pH 9.0. The high ratio of Glc to BSA was chosen to ensure a good degree of irreversible glucosylation . This is because sugars tend to react faster than amino compounds in the Maillard reaction, and they might caramelize, isomerize, fragment, or form diglucosylamines, whereas amino compounds might even be regenerated from the initial condensation product. , From the published literature, it was found that a common ratio for BSA:Glc mixtures is 1:5, which translates to a ratio of 12:1 for glucose per lysine residue.…”

Section: Methodsmentioning

confidence: 99%

Conjugation of Bovine Serum Albumin and Glucose under Combined High Pressure and Heat

Buckow

Wendorff

Hémar

2011

J. Agric. Food Chem.

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The effect of combined heat and pressure on the Maillard reaction between bovine serum albumin (BSA) and glucose was investigated. The effects in the range of 60-132 °C and at 0.1-600 MPa on the lysine availability of BSA were investigated at isothermal/isobaric conditions. The kinetic results showed that the protein-sugar conjugation rate increased with increasing temperature, whereas it decreased with increasing pressure. The reaction followed 1.4th order kinetics at most conditions investigated. A mathematical model describing BSA-glucose conjugation kinetics as a function of pressure and temperature is proposed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-offlight mass spectrometry were used to verify BSA-glucose conjugation and to identify the glucosylated sites. These indicated that the application of combined high pressure and high temperature resulted in significant differences in the progression of the Maillard reaction as compared to heat treatments at atmospheric pressure.

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“…Preparation of glycated BSA Glycated BSA was prepared following the protocol of Kublashvili et al then purified following the protocol of Day (Day et al 1979;Kublashvili and Ugrekhelidze 2005). Because BSA is glycated at the same sites in vitro and in vivo, there was no need to isolate albumin from a live sample (Day et al 1979).…”

Section: Isolation and Purification Of Enterobactinmentioning

confidence: 99%

Enhanced iron availability by protein glycation may explain higher infection rates in diabetics

et al. 2011

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Serum proteins exist in a state of higher glycation among individuals with poor glycemic control, notably diabetics. These non-enzymatic modifications via the Maillard reaction have far reaching effects on metabolism and regulation, and may be responsible for increased infection rates within this population. Here we explore the effects of glycation on iron metabolism and innate immunity by investigating the interaction between siderophores and bovine serum albumin (BSA). Using a quartz crystal microbalance with dissipation monitoring to quantify association rates, glycated BSA exhibited a significantly reduced affinity for apo and holo enterobactin compared to a non-glycated BSA standard. Bacterial growth assays in the presence of BSA and under iron-limited conditions indicated the growth rate of enterobactin-producing E. coli increased significantly when the BSA was in a glycated form. The results, in addition to data in the literature, support the hypothesis that glycation of serum proteins may effectively increase the available free iron pool for bacteria in blood serum and weaken our innate immunity. This phenomenon may be partially responsible for higher infection rates in some diabetics, especially those with poor glycemic control.

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Glycosylation of Bovine Serum Albumin with D-[14C]-Glucose

Abstract: UDC 547.917 Certain parameters of the Maillard reaction between bovine serum albumin (BSA) and D-[ 14 C]-glucose were investigated.

Cited by 3 publications

References 5 publications

Aluminum Oxide Nanoparticles as Carriers and Adjuvants for Eliciting Antibodies from Non-immunogenic Haptens

Aluminum Oxide Nanoparticles as Carriers and Adjuvants for Eliciting Antibodies from Non-immunogenic Haptens

Conjugation of Bovine Serum Albumin and Glucose under Combined High Pressure and Heat

Enhanced iron availability by protein glycation may explain higher infection rates in diabetics

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