A novel expression system of domain I of human beta2 glycoprotein I in -3

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>ntibodies to DI indicates that expressed recombinant DI is likely to be folded correctly

A novel expression system of domain I of human beta2 glycoprotein I in -6

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>d-DI bound to a nickel chelate plate, by recombinant DI and wild type βGPI as inhibitors. A constant amount of antibody was mixed with varying concentrations of inhibitor and allowed to equilibrate at room temperature. For a given μmol concentration of inhibitor, DI alone is a more effective inhibitor of IS4V/IS4Vthan whole βGPI

A novel expression system of domain I of human beta2 glycoprotein I in -4

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>eriplasmic extract; , flow-through from nickel column; , wash with 5 mM imidazole; , wash with 45 mM imidazole; , elution with 300 mM imidazole

A novel expression system of domain I of human beta2 glycoprotein I in -2

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>red onto a nitrocellulose membrane from a 15%-SDS-PAGE gel and probed with an anti-hisantibody. , his-tagged molecular weight protein markers; , DI expressed using the native cDNA human sequence; , DI expressed using sequence optimised for . Equal volumes of periplasmic extract were applied to lanes 1 and 2 on the protein gel

A novel expression system of domain I of human beta2 glycoprotein I in -5

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>. IS4V/IS4Vand IS4V/UK-4V. The same order of binding of aPL to both βGPI and DI was observed. Greatest binding was seen with IS4V/B3Vand least with IS4V/UK-4V

A novel expression system of domain I of human beta2 glycoprotein I in -7

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p> known to bind DI, whole βGPI and CL. IgG was purified from 21 APS patients, 14 SLE patients and 9 normal controls. 20 μg/ml of IgG of each individual patient sample was tested for binding to DI and compared to the standard. Binding to DI was significantly greater in the APS group compared to SLE disease control and normal control groups (Student's t-test analysis (paired, 2-tailed) – APS versus SLE, **p < 0.005; APS versus normal controls, **p < 0.005; SLE versus normal controls p = 0.2)

A novel expression system of domain I of human beta2 glycoprotein I in -0

<p><b>Copyright information:</b></p><p>Taken from "A novel expression system of domain I of human beta2 glycoprotein I in "</p><p>BMC Biotechnology 2006;6():8-8.</p><p>Published online 10 Feb 2006</p><p>PMCID:PMC1402286.</p><p>Copyright © 2006 Ioannou et al; licensee BioMed Central Ltd.</p>er sequence. Codons were chosen for optimal expression of DI in by Juniper, altering 67% of the native sequence (boxed codons). Synthetic oligonucleotide primers are denoted by grey shading; oligonucleotides of the upper DNA sequence are read 5' to 3' (left to right) and the oligonucleotides of the lower DNA sequence are read 3' to 5' (left to right). Restriction sites are denoted in light grey font

Triple, double & single positivity per antibody isotype in APS.

<p>Triple, double & single positivity per antibody isotype in APS.</p

Regression analysis: hazard ratio for APS.

<p>Regression analysis: hazard ratio for APS.</p

ROC analysis: aβ₂GPI tests best discriminate for APS.

<p>Receiver operating characteristic (ROC) analysis was performed to assess the ability of each of the nine aPL assays to discriminate between APS and non-APS subjects. For all three antibody isotypes, the resulting ROC curves illustrate the superiority of aβ₂GPI tests compared to aCL and aDI for APS diagnosis (numerical results are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156407#pone.0156407.t003" target="_blank">Table 3</a>). Abbreviations: GPLU, MPLU, APLU: IgG/IgM/IgA phospholipid units respectively; GBU, MBU, ABU: IgG/IgM/IgA β₂GPI units respectively; GDIU, MDIU, ADIU: IgG/IgM/IgA DI units respectively.</p