Journal: Ultrasound in medicine & biology

Article Title: Accelerated clearance of ultrasound contrast agents containing polyethylene glycol is associated with the generation of anti-polyethylene glycol antibodies

doi: 10.1016/j.ultrasmedbio.2018.02.006

Figure Lengend Snippet: Quantification of serum anti-PEG antibody levels and results of a competitive ELISA using free mPEG-DSPE (experiment 1). (A) Serum anti-PEG IgG is shown to peak 7–14 d after the initial dose of house-MBs and decrease by the 28-d time point. (B) Serum anti-PEG IgM is shown to peak on day 7 for multiple and single dose groups. Anti-PEG IgM was found to decrease at subsequent time points. C and D) For those samples showing high anti-PEG IgM or anti-PEG IgG antibody expression, a competitive ELISA assay was performed to confirm PEG specificity. IgM and IgG binding to the PEG-coated ELISA plate was shown to decrease substantially when serum was incubated in the presence of free mPEG-DSPE (0.9–0.99 mg/ml). * p ≤ 0.05; ** p ≤ 0.01; **** p ≤ 0.0001. ELISA = enzyme-linked immunosorbent assay, Ig = immunoglobulin, DSPE-mPEG2000 = 1,2,-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy(polyethylene-glycol)-2000.

Article Snippet: Serum concentrations of anti-PEG IgM or anti-PEG IgG antibodies were analyzed separately using their respective enzyme-linked immunosorbent assay (ELISA) kits (Life Diagnostics, Inc., West Chester, PA, USA) performed according to the manufacturer’s protocol.

Techniques: Competitive ELISA, Expressing, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Ultrasound in medicine & biology

Article Title: Accelerated clearance of ultrasound contrast agents containing polyethylene glycol is associated with the generation of anti-polyethylene glycol antibodies

doi: 10.1016/j.ultrasmedbio.2018.02.006

Figure Lengend Snippet: Enzyme-linked immunosorbent assay detection of serum anti-PEG IgM (A) and anti-PEG IgG (B) antibodies for animals repeatedly dosed with Definity or Optison over 30 d. PEG = polyethylene glycol; Ig = immunoglobulin

Article Snippet: Serum concentrations of anti-PEG IgM or anti-PEG IgG antibodies were analyzed separately using their respective enzyme-linked immunosorbent assay (ELISA) kits (Life Diagnostics, Inc., West Chester, PA, USA) performed according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Nature Materials

Article Title: STING agonist delivery by tumour-penetrating PEG-lipid nanodiscs primes robust anticancer immunity

doi: 10.1038/s41563-022-01251-z

Figure Lengend Snippet: a-e , Groups of C57Bl/6 mice ( n = 5 animals/group) were treated once by i.v. injection with 5 nmol LND-CDN (red), 5 nmol liposome-CDN (blue), or saline control (black). Shown are serum concentrations of liver enzymes ( a ) alanine aminotransferase, ( b ) aspartate aminotransferase, and ( c ) blood urea nitrogen with the normal ranges indicated by dashed horizontal lines. d , Inflammatory cytokines and chemokines measured by cytokine bead array as a function of time. e , Liver and spleens were collected at 48 hr post dosing for histopathological imaging. Scale bars 100 µm. f , Groups of C57Bl/6 mice ( n = 6 (Non-treated control) or 8 (LND-CDN) animals/group) were inoculated with 3 × 10 5 MC38 tumour cells s.c. in the flank on day 0, then treated on days 7, 14, and 21 with 5 nmol LND-CDN. Serum was collected on day 28 for ELISA analysis of anti-PEG IgG. Shown are raw ELISA absorbances as a function of serum dilution (bottom x-axis) and binding of serial dilutions of a monoclonal anti-PEG antibody standard (‘STD mouse anti-PEG IgG’. top x-axis). Data are shown as mean ± SEM and analysed by two-way ANOVA with Tukey post-test statistical analysis: ns, not significant; *, p < 0.05; **, p < 0.01; ****, p < 0.0001.

Article Snippet: After washing three times with buffer, serum samples and mouse anti-PEG IgG standard antibody (AffinityImmuno kit catalogue number EL-141-PEG-mIGG, starting at 1 μg ml −1 followed by two serial dilutions) were added for 2 h prior to washing.

Techniques: Injection, Saline, Control, Imaging, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Experimental & Molecular Medicine

Article Title: A novel anti-IL4Rα nanoparticle efficiently controls lung inflammation during asthma

doi: 10.1038/emm.2016.89

Figure Lengend Snippet: Proper administration of anti-IL4Rα-NPs to inflamed lung tissue. ( a ) A diagram representing the synthetized anti-IL4Rα nanoparticles. ( b ) Histological assessments confirming the targeting of anti-IL4Rα NPs to the inflamed tissue in asthmatic lungs. (i) H&E staining showing tissue inflammation, (ii) Prussian blue staining (marker for iron oxide) showing distribution of SPION within the inflamed tissue and (iii) immunohistochemical staining for PEG using anti-PEG antibodies also confirmed the presence of SPION. The black arrows indicate the localization of the PEGylated nanoparticles. (iv) Immunofluorescence staining showing the presence of the FITC-anti-IL4Rα-NPs within the tissue. Immunofluorescence images were captured at an exposure of 300 ms. Scale bar, 100 μm.

Article Snippet: Another set of slides were also processed for immunohistochemistry using anti-PEG antibodies (1:200 dilution; BD Pharmingen, Oxford, UK), as previously described.

Techniques: Staining, Marker, Immunohistochemical staining, Immunofluorescence

Journal: Experimental & Molecular Medicine

Article Title: A novel anti-IL4Rα nanoparticle efficiently controls lung inflammation during asthma

doi: 10.1038/emm.2016.89

Figure Lengend Snippet: Treatment with anti-IL4Rα-NPs induces sustained control of lung inflammation. ( a ) Lung sections of OVA-sensitized mice treated with anti-IgG1-NPs, free anti-IL4Rα and anti-IL4Rα-NPs. The sections were isolated 24 h following the last instillation. The upper panels are stained with H&E, the middle panels show Periodic acid Schiff staining (for mucus hypersecretion) and the lower panels show immunohistochemical staining with an anti-PEG antibody to show the tissue distribution of the NPs. ( b ) Histological assessment of lung sections obtained 7 days following the last instillation with either free anti-IL4Rα or anti-IL4Rα-NPs. The upper panels are stained with H&E, and the lower panels show periodic acid-Schiff staining. In contrast to treatment with free anti-IL4Rα, where lung inflammation and obstruction of the bronchiole lumen through mucus accumulation resumed 7 days following the last instillation, lungs treated with anti-IL4Rα-NPs maintained a low level of inflammation and mucus clearance. Scale bar, 100 μm.

Article Snippet: Another set of slides were also processed for immunohistochemistry using anti-PEG antibodies (1:200 dilution; BD Pharmingen, Oxford, UK), as previously described.

Techniques: Isolation, Staining, Immunohistochemical staining