anti caspr2 antibodies  (Thermo Fisher)

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Na V 1.9 labels Domain 2 in calyx endings of dimorphic afferents. (A) Transverse section of a crista showing two dimorphic calyx endings, whose apical parts (Domain 2) are stained for Na V 1.9 (red , arrows) . Inset, left , schematic of entire calyx, showing Domain 2 in red. Inset, right , immunolabeling of Domain 2 with Caspr2 ( green , arrows ) to show similarity to Na V 1.9 immunolabeling. (B) Electron micrograph showing Na V 1.9 label ( arrowheads ) in Domain 2, marked by dashed lines, rather than in the enclosed hair cell or the supporting cells. (C) Surface view of the extrastriolar region of a whole mount utricular macula from an adult female rat. Na V 1.9 antibody ( red circles ) labels the upper portion (Domain 2) of D calyx endings. Most type II hair cells ( II ) and a small proportion of type I hair cells are labeled with calretinin ( solid green , Desai, Zeh, and Lysakowski ). (D, D’) A complex calyx, from the striolar region in the same macula as in A , is labeled with calretinin ( green ), defining it as a C fiber. D is more superficial than D’ . The three calyces in D fuse at deeper levels ( D’ ) to form one triple complex ending. This and other C fibers lacked Na V 1.9 labeling. Scale bars: A, C, D = 5 µm; B = 0.2 µm.

Article Snippet: Caspr2 (ms) , NeuroMab , 75‐075 RRID:AB_2245198 , X (w) , X (Br) , , X , , Datasheet.

Techniques: Staining, Immunolabeling, Labeling

Journal: Translational Psychiatry

Article Title: Heterogeneity of anti-Caspr2 antibodies: specificity and pathogenicity

doi: 10.1038/s41398-025-03677-w

Figure Lengend Snippet: A Diagram of the structure of P13F4 and P6B2 with their unique sequences for the heavy and light chains. B Isotype determination with a sandwich ELISA done by coating a plate with various anti-IgG isotype subtypes, adding purified antibody to each well, and measuring their optical density (OD). P13F4 and P6B2 are IgG2 isotype. C Anti-Casp2 binding on HEK293 cells (top) and GnTI-HEK293 cells (bottom). P13F4 and P6B2 bind preferentially to human Casp2 and less to mouse Caspr2 compared to the null binding of an IgG2-matched isotype control (CON2). D Binding of P13F4 (10 μg/mL) and P6B2 (10 μg/mL) to adult mouse brain sections, PFA-fixed (first column), snap-frozen (second column) and E15.5 whole embryo sections (third column). In brain tissue, P6B2 (second row) binds only to PFA-fixed adult sections, whereas P13F4 (first row) binds to both PFA-fixed adult and E15.5 embryonic sections. IgG2b isotype control third row. Representative images of the CA1 region of the hippocampus (adult, first and second column) and cortical plate (E15.5, third column) at 20x. E Binding of P13F4 (10 μg/mL) and P6B2 (10 μg/mL) to primary neuronal cultures (fixed at DIV14) stained with primary antibodies. Hybridoma anti-Caspr2 antibodies are labeled in green, a commercial anti-Caspr2 antibody (ABN1380) in red, and their co-localization in yellow. Images (P13F4 top, P6B2 middle, IgG2b isotype control bottom) taken at 40x. F Assessment of male mice exposed to anti-Caspr2 antibodies in utero reveals that mice in the P13F4 group have normal behavior in the social interaction task but less time in the center of the chamber during the open field task and more distance covered in the light-dark box task. Mice in the P6B2 group show less body contact with an unfamiliar mouse in the social interaction task. Statistical testing done with ANOVA followed by Tukey post-hoc test. G Sholl analysis of pyramidal neurons from the CA1 and CA2 regions of the hippocampus reveals that P6B2-exposed males have significantly shorter dendrite length when compared to male mice exposed to isotype control in both CA1( P < 0.007, ICC=0.174) and CA2 neurons ( P = 0.0084, ICC = 0.214). Insets (at right in each panel) show representative tracing of CA1 and CA 2 neurons.

Article Snippet: Sections were blocked in 3% BSA in 1x PBS-0.1% Triton X-100 at room temperature for 1 h, incubated with 10 μg/mL of hybridoma generated anti-Caspr2 antibodies overnight at 4 °C, washed with 1x PBS-0.1% Tween-20, and incubated with secondary antibody anti-mouse Alexa Fluor 488 (ThermoFisher Scientific, Cat No A32723, 1:400) for 1 h at room temperature, protected from light.

Techniques: Sandwich ELISA, Purification, Binding Assay, Control, Staining, Labeling, In Utero

Journal: Translational Psychiatry

Article Title: Heterogeneity of anti-Caspr2 antibodies: specificity and pathogenicity

doi: 10.1038/s41398-025-03677-w

Figure Lengend Snippet: A Diagram of the structure of P11G7 and P9C7 with their unique sequences for the heavy and light chains. B Isotype determination with a sandwich ELISA done by coating a plate with various anti-IgG isotype subtypes, adding purified antibody to each well, and measuring their optical density (OD). P11G7 and P9C7 are clearly IgG1 isotype. C Anti-Casp2 binding on HEK293 cells (top) and GnTI-HEK293 cells (bottom). P11G7 and P9C7 bind to both human Casp2 and mouse Caspr2 compared to an IgG1-matched isotype control (CON1). D Binding of P11G7 (10 μg/mL) and P9C7 (10 μg/mL) to adult mouse brain sections, PFA-fixed (first column), snap-frozen (second column) and E15.5 whole embryo sections (third column). P11G7 (top row) binds to all sections, whereas P9C7 (second row) binds to snap-frozen and embryonic sections but not PFA-fixed sections. IgG1 isotype control third row. Representative images of the CA1 region of the hippocampus (adult, first and second column) and cortical plate (E15.5, third column) at 20x. E Binding of P11G7 (10 μg/mL) and P9C7 (10 μg/mL) to primary neuronal cultures (fixed at DIV14) stained with primary antibodies. Hybridoma anti-Caspr2 antibodies are labeled in green, a commercial anti-Caspr2 antibody (ABN1380) in red, and their co-localization in yellow. Images (P11G7 top, P9C7 middle, IgG1 isotype control bottom) taken at 40x. F Assessment of male mice exposed to anti-Caspr2 antibodies in utero reveals that mice in the P11G7 group have no differences in any of the tasks compared to the CON1 group, while mice in the P9C7 group has less body contact with an unfamiliar mouse in the social interaction task, and less time in the center of the chamber in the open field task. Statistical testing done with ANOVA followed by Tukey post-hoc test. ( G ) Sholl analysis of pyramidal neurons from the CA1 and CA2 regions of the hippocampus reveals no significant differences in dendritic length between the groups. Insets (at right in each panel) show representative tracing of CA1 and CA2 neurons.

Article Snippet: Sections were blocked in 3% BSA in 1x PBS-0.1% Triton X-100 at room temperature for 1 h, incubated with 10 μg/mL of hybridoma generated anti-Caspr2 antibodies overnight at 4 °C, washed with 1x PBS-0.1% Tween-20, and incubated with secondary antibody anti-mouse Alexa Fluor 488 (ThermoFisher Scientific, Cat No A32723, 1:400) for 1 h at room temperature, protected from light.

Techniques: Sandwich ELISA, Purification, Binding Assay, Control, Staining, Labeling, In Utero

Journal: Translational Psychiatry

Article Title: Heterogeneity of anti-Caspr2 antibodies: specificity and pathogenicity

doi: 10.1038/s41398-025-03677-w

Figure Lengend Snippet: A Caspr2 mutant constructs. The extracellular portion of Casor2 has 8 domains. The mutant constructs each missing one of the domains were previously generated . Deletions are indicated by the respective amino acid positions (Δaa-aa). The Disc iso construct contains only the discoidin domain. B HEK293T cells were transfected with one of the Caspr2 mutant constructs for 48 h and incubated with each anti-Caspr2 antibody using live staining. P11G7 and P9C7 bind to all mutant constructs except the one missing the discoidin domain. P6B2 and P13F4 do not bind to any of the constructs. Isotype controls showed no binding to any construct. C P11G7 and P9C7 bind to the discoidin domain only construct. P13F4 and P6B2 do not bind to the discoidin domain only construct. D P11G7, P9C7, bind to Caspr2 transfected HEK293T cells. P6B2, P13F4 and the isotype controls do not.

Article Snippet: Sections were blocked in 3% BSA in 1x PBS-0.1% Triton X-100 at room temperature for 1 h, incubated with 10 μg/mL of hybridoma generated anti-Caspr2 antibodies overnight at 4 °C, washed with 1x PBS-0.1% Tween-20, and incubated with secondary antibody anti-mouse Alexa Fluor 488 (ThermoFisher Scientific, Cat No A32723, 1:400) for 1 h at room temperature, protected from light.

Techniques: Mutagenesis, Construct, Generated, Transfection, Incubation, Staining, Binding Assay

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: (A) Exemplary immunocytochemical stainings of CASPR2 (cyan) and patient serum (red) binding to membrane of CASPR2-transfected HEK-293 cells and adult DRG neurons. (B) Stainings for different IgG subclasses (cyan) in 2 exemplary total IgG positive (red) patient sera (P31, P39). (C) Distribution of IgG subclasses (IgG4 only or IgG4 plus additional IgG = IgGX) and pain phenotype with pain—red and orange circles, no pain—light and dark blue circles with circle size related to the number of positive patient sera. (D) Distribution of IgG subclass combinations in patients with pain (red) and without pain (blue). CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia; IgG = immunoglobulin G.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Binding Assay, Membrane, Transfection

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: (A) Microfluidic chamber (MFC) with embryonic DRGs to separate somatic and axonal compartments and investigate VGKC complex organization after CASPR2 aAb incubation in separated compartments. (B) Exemplary pictures using lattice SIM 2 microscopy of soma and axons from DRGs in MFCs treated with healthy control serum. Arrows mark the distance between CASPR2 (magenta) and Kv1.2 (green). (C) Quantification of distance between CASPR2 and Kv1.2 for somata (left) and axons (right) after CASPR2 aAb incubation for 2 hours. Data are shown as mean ± SEM. Somatic and axonal complexes: n = 325–1,465 and n = 1872–3,603, respectively. (D) CASPR2 and Kv1.2 density per 100 µm axon (CASPR2 n = 34–37; Kv1.2 n = 33–37). Data are shown as violin plots with individual values, median = bold line, quartiles = dotted lines. Levels of significance: ns: not significant, * p < 0.05, ** p < 0.01, **** p < 0.0001. CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia; VGKC complex = voltage-gated potassium channel complex.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Incubation, Microscopy, Control

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: (A) Exemplary pictures and activity graphs of ROIs after short-term incubation with control serum or serum group with pain and IgGX. Neuronal activity from ∼40 cells is shown in a heatmap. (B) Frequency (left), amplitude (middle), and area under the curve (AUC; right) of spontaneous calcium activity events after short-term incubation with CASPR2 aAbs of different serum subclassifications. Data shown as mean ± SEM, n = 718 (control), n = 682 (no pain/IgG4), n = 476 (no pain/IgGX), n = 566 (pain/IgG4), and n = 727 (pain/IgGX). Levels of significance: * p < 0.05, ** p < 0.01, **** p < 0.0001. aAbs = autoantibodies; CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Activity Assay, Incubation, Control

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: (A) Bar plot of potassium channel activity at different voltage steps from −80 to +40 mV. Normalized current compared with the healthy control is shown. (B) I–V (current-voltage relation) plot measured after short-term aAb application on DRG neurons under the condition: pain/no IgG4, N = 2, n = 10. (C and D) Replacement of the CASPR2 aAbs by healthy control serum led to a rescue of affected potassium channel current (patients with pain and patients without pain), N = 2, n = 10–11. Levels of significance: * p < 0.05, ** p < 0.01, *** p < 0.001. aAbs = autoantibodies; CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia; Kv = voltage-gated potassium.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Activity Assay, Control

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: (A and B) Representative potassium channel current recording and the effect of specific potassium channel subunit composition blocker α-dendrotoxin Dtx-A and κM-RIIIJ (red traces) shown in the bar plot (at +40 mV). Data shown as mean ± SEM. (C) Domain architecture of CASPR2 including the discoidin domain, fibrinogen-like domain, and laminin domains. Patch clamp recordings using a voltage step protocol from −80 to +40 mV in the presence of CASPR2 monoclonal autoantibodies against the discoidin domain (left) demonstrating significant alterations of the potassium channel activity and against the laminin domain (right) with no significant alterations. N = 3, n = 13–16. Level of significance: * p < 0.05, **** p < 0.0001. CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Patch Clamp, Activity Assay

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: Neuropathic Pain and Distinct CASPR2 Autoantibody IgG Subclasses Drive Neuronal Hyperexcitability

doi: 10.1212/NXI.0000000000200423

Figure Lengend Snippet: DRG neurons are active mediators in developing neuropathic pain: healthy condition (left) and disease condition in the presence of CASPR2 autoantibodies (aAbs, right). Sensory neuron hyperexcitability is driven by pain-associated CASPR2 aAbs. This causes enhanced neuronal activity and decreased function of the associated Kv channels. Pathologic activity of sensory neurons is mainly promoted by CASPR2 aAbs of the IgG4 subtype. Created in BioRender. Villmann, C. (2025) BioRender.com/ . CASPR2 = Contactin-associated protein 2; DRG = dorsal root ganglia.

Article Snippet: For live staining, HEK-293 cells 72 hours posttransfection (eMethods) or adult DRG neurons at DIV3 were incubated with anti-CASPR2 antibodies (AF5145, 1:250, R&D Systems, Minneapolis, MN, US) and/or human patient serum (1:50–1:250) in medium for 1 hour at 4°C.

Techniques: Activity Assay

Journal: Science Advances

Article Title: Permissive central tolerance plus defective peripheral checkpoints license pathogenic memory B cells in CASPR2-antibody encephalitis

doi: 10.1126/sciadv.adr9986

Figure Lengend Snippet: ( A ) Left: Representative flow cytometry cell gating strategy to isolate MBCs (red) and NBCs (blue) for bulk (top) and single (bottom, dotted line) B cell cultures. Representative fluorescence microscopy images of culture supernatant detection of secreted CASPR2-reactive IgG or IgM using a live cell–based assay with CASPR2–enhanced green fluorescent protein (EGFP) expressing HEK293T cells. 4′,6-Diamidino-2-phenylindole (DAPI), nuclear stain. Scale bar, 10 μm. Right: mRNA was extracted from CASPR2-reactive single B cell cultures to amplify and clone paired heavy- and light-chain BCR sequences. These were expressed in HEK293S cells to secrete CASPR2-reactive IgG (blue) or IgM (pink). ( B ) The proportion of bulk B cell culture wells containing CASPR2-reactive IgM or IgG from patients (CASPR2-Ab-E, n = 6; LGI1-Ab-E, n = 4; NMDAR-Ab-E, n = 4) and HCs ( n = 5). Black bars depict the median value. ( C ) Donut plots visualizing the frequency and clonality of CASPR2-reactive BCRs in single-cell cultures. Numerator, total number of CASPR2-reactive BCRs; denominator, total number of cells screened. The absolute percentage of CASPR2-reactive BCRs is then shown for two patients (P1 and P2) and two HCs (H5 and H6).

Article Snippet: Commercial polyclonal rabbit anti-human CASPR2 antibody (Novus, A45565) was used as an isotype control for detecting denatured CASPR2.

Techniques: Flow Cytometry, Fluorescence, Microscopy, Cell Based Assay, Expressing, Staining, Cell Culture

Journal: Science Advances

Article Title: Permissive central tolerance plus defective peripheral checkpoints license pathogenic memory B cells in CASPR2-antibody encephalitis

doi: 10.1126/sciadv.adr9986

Figure Lengend Snippet: ( A ) Ig heavy- and light-chain variable region mutation counts across B cell subsets in both patients (CASPR2-Ab-E) and HCs. ( B ) Raw surface plasmon resonance (SPR) traces representing the soluble extracellular domain of human CASPR2 binding to immobilized CASPR2 memory mAbs (top row) and their corresponding UCAs (middle row). E06 UCA mAb did not express. UCA binding as an IgM was tested via a live cell–based assay with “+” indicating CASPR2 reactivity (bottom row). NA, not applicable. ( C ) K D (M) quantification of mAbs via SPR (left). Nonsignificant K D Pearson’s correlations with heavy (middle)– and light (right)–chain mutation count.

Article Snippet: Commercial polyclonal rabbit anti-human CASPR2 antibody (Novus, A45565) was used as an isotype control for detecting denatured CASPR2.

Techniques: Mutagenesis, SPR Assay, Binding Assay, Cell Based Assay

Journal: Science Advances

Article Title: Permissive central tolerance plus defective peripheral checkpoints license pathogenic memory B cells in CASPR2-antibody encephalitis

doi: 10.1126/sciadv.adr9986

Figure Lengend Snippet: ( A ) No differences in the number of CASPR2 peptides after immunoprecipitation (IP) by peptide phage display versus isotype control mAb. ( B ) Representative immunohistochemistry staining (all inlays = isotype control mAb) of CASPR2 mAbs on fixed murine hippocampal brain tissue with hippocampus visualized (top left; mAb, E07) in wild-type (WT) and CASPR2 knockout (−/−; top middle) tissue. Representative immunofluorescent staining using E08 on live hippocampal neurons (top right) fixed rat cerebellum (lower left; DAPI, nuclear counterstain), live human iPSC–derived sensory neurons (bottom middle; mAb, E08), and live mouse dorsal root ganglia (bottom right; mAb, E08). Costaining markers to identify cell types include microtubule-associated protein 2 (MAP2) and β-tubulin III. Scale bars, 500 μm (brain tissue) and 10 μm (all others). ( C ) Endpoint titrations (1:dilutions) of binding across human and mouse CASPR2 live cell–based assays. mAbs are colored as in (F). ( D ) Cartoon representation of CASPR4 single domains knocked into full-length human CASPR2-EGFP (top). Heatmap depicts CASPR4 knock-in domains that abrogated mAb binding (bottom). FibC, Fibrinogen C. ( E ) Discoidin (Disc) and laminin3 (L3) domain amino acid sequences from human and mouse, showing nonconserved amino acids in green. The tan and pink highlight Disc and L3 domains throughout the figure, respectively. Predicted CASPR2 protein structure (right; AlphaFold ID 5Y4M) showing nonconserved amino acids (green), Disc, and L3 domains. ( F ) Binding competition map (left) demonstrating the displacement of a prebound mAb ( x axis) by a competing mAb preconjugated with Alexa Fluor 594 (AF594; y axis). Percentage inhibition is defined as percentage reduction of fluorescence intensity of the respective mAb compared to isotype control and represented by a forced directed network of epitope binning (right). Arrowhead indicates the direction of binding competition and line thickness, and intensity denotes percentage inhibition. Shapes denote patient sample.

Article Snippet: Commercial polyclonal rabbit anti-human CASPR2 antibody (Novus, A45565) was used as an isotype control for detecting denatured CASPR2.

Techniques: Immunoprecipitation, Control, Immunohistochemistry, Staining, Knock-Out, Derivative Assay, Binding Assay, Knock-In, Inhibition, Fluorescence

Journal: Science Advances

Article Title: Permissive central tolerance plus defective peripheral checkpoints license pathogenic memory B cells in CASPR2-antibody encephalitis

doi: 10.1126/sciadv.adr9986

Figure Lengend Snippet: ( A ) Representative images used to calculate colocalization of pHrodo-conjugated IgG4 mAbs and CASPR2-EGFP, reflecting receptor internalization by CASPR2-expressing HEK293T cells (left). Internalization quantified by mean pHrodo fluorescence intensity mAbs in the presence and absence of dynamin inhibition (dynasore; right). ( B ) Pearson’s correlation of receptor internalization to mAb binding parameters; bottom x -axis label denotes internalization. Background colors indicate the three epitope pockets as presented in ; top x -axis label annotates epitope. ( C ) Representative images of CASPR2 and MAP2 expression after a 4-hour application of IgG1 (open circles, left) or IgG4 mAbs (closed circles). Scale bars, 5μm. CASPR2 puncta intensity summarized on the right with * P < 0.05, ** P < 0.01, nonparametric Kruskal-Wallis with Dunn’s multiple comparisons post hoc test. mAbs colored as in (A). ( D ) Representative images (left) and quantification (right) of synaptic AMPAR and PSD95 expression similar to (C). ( E ) Representative tracings (top), single average event and cumulative probability (middle), and amplitude quantifications (bottom) of AMPAR-mediated mEPSC recordings of pyramidal neuronal cultures. * P < 0.05, *** P < 0.001, Kruskal-Wallis test with Dunn’s multiple comparisons post hoc test. ( F ) Cartoon depiction of intracerebral mAb injection into bilateral hippocampal CA3 regions (left). Open-field (middle) and light-dark box (right) behavioral test performance was assessed at 6 and 9 hours postinjection, respectively. * P < 0.05, t test. ( G ) Summary model cartoon.

Article Snippet: Commercial polyclonal rabbit anti-human CASPR2 antibody (Novus, A45565) was used as an isotype control for detecting denatured CASPR2.

Techniques: Expressing, Fluorescence, Inhibition, Binding Assay, Injection

Journal: Science Advances

Article Title: Permissive central tolerance plus defective peripheral checkpoints license pathogenic memory B cells in CASPR2-antibody encephalitis

doi: 10.1126/sciadv.adr9986

Figure Lengend Snippet: ( A ) Heatmaps depicting binding by CASPR2-reactive fab segments, -IgG or -IgM; red, positive binding by live cell based assay (LCA). * P < 0.05 for fab and IgG frequency distributions. Populations labeled in green depict HC, and populations labeled in red depict CASPR2-Ab-E throughout the figure. ( B ) CASPR2-IgM mAb endpoint dilution (EPD) in a live cell–based assay. All mAbs expressed as class IgM to prevent class confound; * P < 0.05 and ** P < 0.005. ( C ) Quantification of the number of peptides enriched in phage immunoprecipitation; **** P < 0.0001, Wilcoxon unpaired. ( D ) Summary cartoon model.

Article Snippet: Commercial polyclonal rabbit anti-human CASPR2 antibody (Novus, A45565) was used as an isotype control for detecting denatured CASPR2.

Techniques: Binding Assay, Cell Based Assay, Labeling, Immunoprecipitation