mouse anti-tpp1 monoclonal antibody  (BioMarin Inc)

Journal: Developmental cell

Article Title: NPC1-mTORC1 signaling Couples Cholesterol Sensing to Organelle Homeostasis and is a Targetable Pathway in Niemann-Pick type C

doi: 10.1016/j.devcel.2020.11.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal anti-CLN2 (TPP1) , Santa Cruz Biotechnology , Cat#sc-393961.

Techniques: Labeling, Plasmid Preparation, Recombinant, Control, Software, Magnetic Beads

Journal: Science Advances

Article Title: Exploiting the diphtheria toxin internalization receptor enhances delivery of proteins to lysosomes for enzyme replacement therapy

doi: 10.1126/sciadv.abb0385

Figure Lengend Snippet: ( A ) Design of LTM-TPP1 fusion protein and delivery schematic. ( B ) Enzyme kinetics of rTPP1 and LTM-TPP1 against the synthetic substrate AAF-AMC are indistinguishable. Michaelis-Menten plots were generated by varying [AAF-AMC] at a constant concentration of 10 nM enzyme (means ± SD; n = 3). Plots and kinetic parameters were calculated with GraphPad Prism 7.04. ( C ) Maturation of TPP1 is unaffected by the N-terminal fusion of LTM. ( D ) LTM-TPP1 inhibits wild-type DT activity in a dose-dependent manner (IC 50 of 17.2 nM), while rTPP1 has no effect on protein synthesis inhibition by DT (means ± SD; n = 3). ( E ) LTM and DTR-TPP1 bind HBEGF with apparent K d ’s of 13.3 and 19.1 nM, respectively. ( F ) LTM-TPP1 colocalizes with LAMP1 staining (red).

Article Snippet: Membranes were then blocked for 1 hour with a 5% milk–tris-buffered saline (TBS) solution and incubated overnight at room temperature with a 1:100 dilution of mouse monoclonal antibody against TPP1 (Abcam, ab54685) in 5% milk-TBS.

Techniques: Generated, Concentration Assay, Activity Assay, Inhibition, Staining

Journal: Science Advances

Article Title: Exploiting the diphtheria toxin internalization receptor enhances delivery of proteins to lysosomes for enzyme replacement therapy

doi: 10.1126/sciadv.abb0385

Figure Lengend Snippet: ( A ) CLN2 knockout cells exhibit ~4% TPP1 activity relative to wild-type HeLa Kyoto cells (means ± SD; n = 3). ( B ) Western blotting against TPP1 reveals no detectable protein in the knockout cells. ( C ) (Left) In vitro maturation of pro-rTPP1 and LTM-TPP1 (16 ng) was analyzed by Western blot. (Right) TPP1 present in wild-type (WT) and TPP1 −/− cells, and TPP1 −/− cells treated with 100 nM rTPP1 and LTM-TPP1. ( D ) Uptake of rTPP1 and LTM-TPP1 into HeLa Kyoto TPP1 −/− cells was monitored by TPP1 activity (means ± SD; n = 4). ( E ) TPP1 activity present in HeLa Kyoto TPP1 −/− cells following a single treatment with 50 nM LTM-TPP1 (means ± SD; n = 3).

Article Snippet: Membranes were then blocked for 1 hour with a 5% milk–tris-buffered saline (TBS) solution and incubated overnight at room temperature with a 1:100 dilution of mouse monoclonal antibody against TPP1 (Abcam, ab54685) in 5% milk-TBS.

Techniques: Knock-Out, Activity Assay, Western Blot, In Vitro

Journal: Science Advances

Article Title: Exploiting the diphtheria toxin internalization receptor enhances delivery of proteins to lysosomes for enzyme replacement therapy

doi: 10.1126/sciadv.abb0385

Figure Lengend Snippet: Uptake of rTPP1 and LTM-TPP1 into HeLa Kyoto TPP1 −/− cells following EndoH treatment to remove N -glycan labeling (means ± SD; n = 3).

Article Snippet: Membranes were then blocked for 1 hour with a 5% milk–tris-buffered saline (TBS) solution and incubated overnight at room temperature with a 1:100 dilution of mouse monoclonal antibody against TPP1 (Abcam, ab54685) in 5% milk-TBS.

Techniques: Glycoproteomics, Labeling

Journal: Science Advances

Article Title: Exploiting the diphtheria toxin internalization receptor enhances delivery of proteins to lysosomes for enzyme replacement therapy

doi: 10.1126/sciadv.abb0385

Figure Lengend Snippet: ( A ) Assay schematic. ( B ) TPP1 activity in brain homogenates of 6-week-old mice injected with two doses (5 and 25 μg) of either rTPP1 or LTM-TPP1 (5 μg, P = 0.01; 25 μg, P = 0.002). ( C ) TPP1 activity in brain homogenates following a single 25-μg dose of LTM-TPP1, 1, 7, and 14 days postinjection. Data are presented as box and whisker plots, with whiskers representing minimum and maximum values from n ≥ 4 mice per group. Statistical significance was calculated using paired t tests with GraphPad Prism 7.04.

Article Snippet: Membranes were then blocked for 1 hour with a 5% milk–tris-buffered saline (TBS) solution and incubated overnight at room temperature with a 1:100 dilution of mouse monoclonal antibody against TPP1 (Abcam, ab54685) in 5% milk-TBS.

Techniques: Activity Assay, Injection, Whisker Assay

Journal: Experimental eye research

Article Title: Intracerebroventricular gene therapy that delays neurological disease progression is associated with selective preservation of retinal ganglion cells in a canine model of CLN2 disease

doi: 10.1016/j.exer.2016.03.023

Figure Lengend Snippet: ERG b-wave amplitudes versus age in CLN2-affected dogs treated with CNS gene therapy (n = 2) compared with those of normal dogs (n = 7) and those of untreated, CLN2-affected dogs (n = 5); control data adapted from . Amplitudes reflect scotopic mixed rod and cone responses after high intensity light flashes of 13.2 log photons/cm 2 /s (10 cd.s/m 2 ). Amplitudes from treated affected dogs were significantly reduced from normal by 6 months of age and continued to decline over time until 12 months of age when no response was detected even at the highest stimulus intensity (*p < 0.05; *p < 0.005). There were no significant differences between treated and untreated CLN2-affected dogs for any of the ages tested. Error bars represent standard deviation.

Article Snippet: Sections of the paraffin-embedded retinas were immunostained with a mouse monoclonal anti-TPP1 antibody diluted 1:100 (Abcam ab54685).

Techniques: Control, Standard Deviation

Journal: Experimental eye research

Article Title: Intracerebroventricular gene therapy that delays neurological disease progression is associated with selective preservation of retinal ganglion cells in a canine model of CLN2 disease

doi: 10.1016/j.exer.2016.03.023

Figure Lengend Snippet: Total retinal thickness decreased progressively during the extended lifespan beyond 12 months of age in gene therapy treated CLN2-affected dogs. While superior and inferior retinal thickness appears be reduced from normal at 14 months, a statistically significant difference from the normal dogs could not be detected at this age due to the small sample sizes. Thickness measurements were taken 1 optic disc diameter (OD) superior to the ONH and 1 OD inferior to the ONH for each timepoint. Error bars represent standard error of the mean (SEM).

Article Snippet: Sections of the paraffin-embedded retinas were immunostained with a mouse monoclonal anti-TPP1 antibody diluted 1:100 (Abcam ab54685).

Techniques:

Journal: Experimental eye research

Article Title: Intracerebroventricular gene therapy that delays neurological disease progression is associated with selective preservation of retinal ganglion cells in a canine model of CLN2 disease

doi: 10.1016/j.exer.2016.03.023

Figure Lengend Snippet: OCT cross-sectional images showing thinning of the superior and inferior retina in CLN2-affected dog B at 17 months of age. OCT images from a normal dog of similar age, taken from comparable locations in the retina, are included for comparison.

Article Snippet: Sections of the paraffin-embedded retinas were immunostained with a mouse monoclonal anti-TPP1 antibody diluted 1:100 (Abcam ab54685).

Techniques: Comparison

Journal: Experimental eye research

Article Title: Intracerebroventricular gene therapy that delays neurological disease progression is associated with selective preservation of retinal ganglion cells in a canine model of CLN2 disease

doi: 10.1016/j.exer.2016.03.023

Figure Lengend Snippet: Numbers of axons in optic nerve cross sections of 3 TPP 1−/− dogs that received intracerbroventricular TPP1 gene therapy.

Article Snippet: Sections of the paraffin-embedded retinas were immunostained with a mouse monoclonal anti-TPP1 antibody diluted 1:100 (Abcam ab54685).

Techniques:

Journal: Experimental eye research

Article Title: Intracerebroventricular gene therapy that delays neurological disease progression is associated with selective preservation of retinal ganglion cells in a canine model of CLN2 disease

doi: 10.1016/j.exer.2016.03.023

Figure Lengend Snippet: Light micrographs of retinal cross-sections immunostained with an anti-TPP1 antibody. The brown color indicates TPP1 antibody binding. (A) The retina from a normal Dachshund shows punctate immunostaining in the ganglion cells (arrows in A). (B) Ganglion cells from Dog B did not show any TPP1 immunolabeling (arrow in B). Intense TPP1 immunostaining was also present around the photoreceptor outer segments in the normal dog and diffuse immunostaining was present throughout most of the rest of the retina. Both retinas were artifactually detached from the retinal pigment epithelium during processing for paraffin embedding. Bar in (B) indicates the magnification of both micrographs.

Article Snippet: Sections of the paraffin-embedded retinas were immunostained with a mouse monoclonal anti-TPP1 antibody diluted 1:100 (Abcam ab54685).

Techniques: Binding Assay, Immunostaining, Immunolabeling