Optimization of Unnicked<i>β</i>2-Glycoprotein I and High Avidity Anti-<i>β</i>2-Glycoprotein I Antibodies Isolation

Artenjak, Andrej; Leonardi, Adrijana; Križaj, Igor; Ambrožič, Aleš; Sodin‐Šemrl, Snežna; Božič, Borut; Čučnik, Saša

doi:10.1155/2014/195687

Cited by 7 publications

(4 citation statements)

References 29 publications

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“…β2GPI was isolated from human plasma using a combination of perchloric acid precipitation and heparin affinity/cationic exchange chromatography. 61 Human polyclonal IgG were isolated from sera of healthy blood donors by affinity chromatography HiTrap™ Protein G HP column (GE HealthCare Bio-Sciences AB, Uppsala, Sweden) as previously described. Oxidatively altered IgG (oxIgG) with increased immunoreactivity to β2GPI and its peptide clusters influence human coronary artery endothelial cells.…”

Section: Biological Materialsmentioning

confidence: 99%

Binding of plasma proteins to titanium dioxide nanotubes with different diameters

Kulkarni

Flašker

Lokar

et al. 2015

IJN

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Titanium and titanium alloys are considered to be one of the most applicable materials in medical devices because of their suitable properties, most importantly high corrosion resistance and the specific combination of strength with biocompatibility. In order to improve the biocompatibility of titanium surfaces, the current report initially focuses on specifying the topography of titanium dioxide (TiO 2 ) nanotubes (NTs) by electrochemical anodization. The zeta potential (ζ-potential) of NTs showed a negative value and confirmed the agreement between the measured and theoretically predicted dependence of ζ-potential on salt concentration, whereby the absolute value of ζ-potential diminished with increasing salt concentrations. We investigated binding of various plasma proteins with different sizes and charges using the bicinchoninic acid assay and immunofluorescence microscopy. Results showed effective and comparatively higher protein binding to NTs with 100 nm diameters (compared to 50 or 15 nm). We also showed a dose-dependent effect of serum amyloid A protein binding to NTs. These results and theoretical calculations of total available surface area for binding of proteins indicate that the largest surface area (also considering the NT lengths) is available for 100 nm NTs, with decreasing surface area for 50 and 15 nm NTs. These current investigations will have an impact on increasing the binding ability of biomedical devices in the body leading to increased durability of biomedical devices.

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Section: Biological Materialsmentioning

confidence: 99%

Binding of plasma proteins to titanium dioxide nanotubes with different diameters

Kulkarni

Flašker

Lokar

et al. 2015

IJN

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“…All experiments were performed at 25 C in a 1 cm path length disposable low-volume cuvette containing 100 ml of the sample with concentrations 1.3 g/l of b2GPI and 0.75 g/l of IgG6 and HAv IgG anti-b2GPI, respectively. As an antigen and antibody protein sample, aliquots of b2GPI and HAv IgG anti-b2GPI, isolated and purified in our previous research studies, 22,33 were used. Antigen-antibody interactions were determined by mixing 50 ml of antibody solution with 50 ml of antigen b2GPI solution and making rapid sizing measurements (every 30 seconds for 60 minutes).…”

Section: Dynamic Light-scattering (Dls) Analysismentioning

confidence: 99%

Oxidatively altered IgG with increased immunoreactivity to β2-glycoprotein I and its peptide clusters influence human coronary artery endothelial cells

Artenjak

Omersel

Grabnar

et al. 2015

Lupus

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Oxidative stress has been shown to play a role in modifying antibodies in favor of higher auto-immunoreactivity. We studied the immunoreactivity of oxidized IgG (oxIgG) to β2-glycoprotein I (β2GPI), six peptide sequences corresponding to amino acid clusters on its different domains, to determine their effects on human coronary artery endothelial cells (HCAEC). Human IgG was purified from seven donors, electro-oxidized and checked for immunoreactivity and avidity to β2GPI and to peptides by ELISA. Conformational stability and antibody-antigen complex formation of oxIgG was analyzed by fluorescence spectroscopy and dynamic light scattering. Resting and activated sub-confluent HCAEC were stimulated with oxIgG or IgG. Secreted cytokines were measured by ELISA. Immunoreactivity of seven oxIgG samples increased to 7.5-fold against β2GPI and to 3.8-fold against six peptides as compared to IgG. oxIgG showed low avidity "properties." Conformational changes and exposure of protein hydrophobic regions were confirmed by an elevation in fluorescence (2.4- to 5.0-fold) on bis-ANS dye binding to oxIgG. oxIgG significantly elevated the release of GROα and IL-8 in resting and activated states of HCAEC. Oxidation alters IgG in favor of autoreactivity toward whole β2GPI and corresponding peptides on different domains of β2GPI and could lead to dysfunction of arterial endothelium by upregulation of chemokines.

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“…ELISA that measures the immune reactivity directly to β 2 GPI is used for the detection of antiβ 2 GPI (IgG and IgM). The characteristics are similar to the detection of aCL with the major difference that in anti-β 2 GPI ELISA, the purified human native β 2 GPI [94] is coated directly onto the ELISA microtitre plates. Briefly, high binding polystyrene microtitre plates were coated with β 2 GPI in PBS and incubated with sera diluted in PBS containing 0.05% Tween-20.…”

Section: Antibodies Against β 2 Gpimentioning

confidence: 98%

Laboratory Methodology Important in the Diagnosis and Prognosis of Antiphospholipid Syndrome

Žigon¹,

Mijovski²,

Frank-Bertoncelj³

et al. 2015

Thrombosis, Atherosclerosis and Atherothrombosis - New Insights and Experimental Protocols

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Antiphospholipid syndrome (APS) is an autoimmune disease, characterized by thrombosis and pregnancy complications with persistently elevated levels of antiphospholipid antibodies (aPL). Recently, a unique mathematical calculation has been presented to assess the risk of thrombosis in patients with APS called antiphospholipid score or global antiphospholipid syndrome score (GAPSS). This new approach in the diagnosis of APS leads to the assessment of the risk of thrombosis considering the results of different aPL (lupus anticoagulants (LA), anticardiolipin antibodies (aCL), antibodies against β2GPI (anti-β2GPI), and phosphatidylserinedependent antiprothrombin antibodies (aPS/PT) (isotypes IgG and IgM). This chapter provides an overview of the algorithm strategy for APS diagnosis with the aims of characterizing in detail the laboratory methodology of criteria aPL (LA, aCL, and anti-β2GPI) and noncriteria aPL, such as IgA aCL and IgA anti-β2GPI, anti-domain I β2GPI, and antiprothrombin antibodies. In order to improve APS diagnosis, several new approaches in aPL detection have recently been suggested, such as multiline immunodot assay, detection of aPL by flow cytometry using beads with particular surface properties, and the newly developed automated BioPlex system technology for parallel detection of aCL and anti-β2GPI antibodies of IgG, IgA, and IgM isotypes. A completely different and promising approach in future research lies in the potential of microRNAs as biomarkers for risk of thrombosis and/or obstetric complication.

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Optimization of Unnickedβ2-Glycoprotein I and High Avidity Anti-β2-Glycoprotein I Antibodies Isolation

Cited by 7 publications

References 29 publications

Binding of plasma proteins to titanium dioxide nanotubes with different diameters

Binding of plasma proteins to titanium dioxide nanotubes with different diameters

Oxidatively altered IgG with increased immunoreactivity to β2-glycoprotein I and its peptide clusters influence human coronary artery endothelial cells

Laboratory Methodology Important in the Diagnosis and Prognosis of Antiphospholipid Syndrome

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