Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Evaluation of the cytotoxic activity of NK cells treated with anti-PSMA Ab in vitro . (A, B) The killing rates of NK cells against 22RV1 cells (PSMA strongly positive) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the Cell Counting Kit-8 (CCK-8) assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (A) , 1:1 (B) , respectively; (C, D) The killing rates of NK cells against PC3 cells (PSMA negative) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the CCK-8 assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (C) , 1:1 (D) , respectively; (E, F) The killing rates of NK cells against RWPE-1 cells (PSMA moderately positive) in the NK, NK + IgG (10 μg/mL), NK + anti-PSMA Ab (5, 10, 20 μg/mL) treatment groups measured using the CCK-8 assay at 2 and 6 h after co-culturing (n = 3). E/T = 0.5:1 (E) , 1:1 (F) , respectively; (G) PSA levels in the culture supernatant of NK cells co-cultured with 22RV1 cells in the control and treatment groups, including NK, NK + IgG, and NK + anti-PSMA Ab (10 μg/mL), measured via ELISA at 2, 6, 12, and 24 h after co-culturing (n = 3); (H) Representative flow cytometry plots and summary data (n = 3) of the MFI for CD107a expression in NK cells co-cultured with 22RV1 cells in the presence of anti-PSMA Ab (10 μg/mL) or IgG control (10 μg/mL). CD107a expression in NK cells was set as a negative control. Degranulation of NK cells was induced upon interaction with 22RV1 cells at a 1:1 ratio with anti-PSMA Ab or IgG for 6 h at 37 °C, the GolgiStop protein transport inhibitor was added during the final 2 h of the culture and NK cells were collected for cytometry measurement; (I, J) Comparison of supernatant perforin (I) and granzyme B (J) levels among NK cells co-cultured with 22RV1 cells and their counterparts co-cultured with 22RV1 cells in the presence of IgG control (10 μg/mL) or anti-PSMA Ab (10 μg/mL) at E:T of 1:1 after 6h coculture (n = 3). NK cells alone were set as negative control; (K, L) Comparison of IFN-γ (K) and TNF-α (L) levels among NK cells co-cultured with 22RV1 cells and their counterparts co-cultured with 22RV1 cells in the presence of IgG control (10 μg/mL) or anti-PSMA Ab (10 μg/mL) at E:T of 1:1 after 6h co-culture (n=3). NK cells alone were set as negative control; Data expressed as means ± SD were plotted, and ANOVA followed by a Tukey’s post hoc test was used to compare three or more groups (A–L) . *p < 0.05; ns, not significant. anti-PSMA Ab, anti-prostate-specific membrane antigen antibody; PSA, prostate-specific antigen; ELISA, enzyme-linked immunosorbent assay; IFN-γ, interferon-γ; TNF-α, tumor necrosis factor-α; E:T, effector-to-target ratio.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: Activity Assay, In Vitro, Cell Counting, CCK-8 Assay, Cell Culture, Control, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Negative Control, Cytometry, Comparison, Co-Culture Assay, Membrane

Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Development of PDO PCa models and cytotoxicity of combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells against the PDO. (A) Representative hematoxylin-and-eosin images of PCa tissue-derived organoid derived from PCa specimens (scale bar 200 μm). The left image was the panorama, and the right four images were local magnifications part by part (a→a’, b→b’, c→c’, d→d’); (B) Representative PSMA immunohistochemistry images of PCa tissue-derived organoid derived from PCa specimens (scale bar 200 μm). The left image was the panorama, and the right four images were local magnifications part by part (a→a’, b→b’, c→c’, d→d’); (C) Representative bright-field image of coculture of NK cells with PCa tissue-derived organoid in the presence of IgG or the constructed anti-PSMA antibody after 2 h and 6h coculture, PDOs alone were set as controls (n = 3); (D) Cytotoxicity of NK cells with IgG or anti-PSMA antibody (10 μg/mL) against PCa tissue-derived organoid at E/T ratio of 5:1 after 2 h and 6h coculture measured using LDH assay (n = 3); (E) IFN-γ levels of the supernatant after the NK cells were co-cultured with PCa tissue-derived organoid at E/T ratio of 5:1 after 2 h and 6h coculture measured using ELISA (n = 3). Data are shown as mean ± SD. Statistical significance was determined using an unpaired t-test (D, E) . *p < 0.05; ns, not significant. PDO, patient-derived organoid; PCa, prostate cancer; PSMA, prostate-specific membrane antigen; IgG, immunoglobulin G; E/T, effector-to-target ratio; LDH, lactate dehydrogenase; IFN-γ, interferon-gamma; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: Derivative Assay, Immunohistochemistry, Construct, Lactate Dehydrogenase Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Membrane

Journal: Frontiers in Immunology

Article Title: Combined treatment with anti-PSMA antibody and human peripheral blood-derived NK cells for castration-resistant prostate cancer

doi: 10.3389/fimmu.2025.1572676

Figure Lengend Snippet: Anti-tumor effect of NK cells against CRPC in combination with anti-PSMA Ab in a subcutaneous tumor model in vivo . (A) Experimental protocol for the CRPC model used in (B–I) : mice were injected with PBS, anti-PSMA Ab or isotype-matched control mAb (10 mg/mg) intraperitoneally (i.p.) on days 10, 18 and injected with NK cells (1 × 10 7 ) intravenously (i.v.) on days 11, 15, 19, and 23 after injections of 2 × 10 6 22RV1 cancer cells subcutaneously (s.c.) on day 0 (n = 6 per group); (B) Tumor volumes at various times (horizontal axis) after tumor inoculation in the control, NK, NK + IgG, and NK + anti-PSMA Ab groups. Tumor volumes were calculated according to the formula L × W 2 /2, where L and W represent the longest and shortest diameters measured using a caliper, respectively (n = 6 per group); (C) Body weights in the control and treatment groups over the whole treatment course (n = 6 per group); (D) Serum PSA levels of mice in the control, NK, NK + IgG, and anti-PSMA Ab groups (n = 6 per group) on days 10, 16, 22, and 28; (E) Images of tumors in mice 28 days after tumor inoculation (n = 5–6 in each group); (F) Tumor weights corresponding to each group when harvested on day 28 (n = 5–6 in each group); (G) HE examination of tumor specimen in the control and treatment groups on day 28 (n = 5–6 in each group); (H) Serum IL-6 levels in the control and treatment groups on day 28; (I) Cumulative Kaplan–Meier survival curves for mice (n = 6 per group) after tumor implantation. Data expressed as the means ± SD were plotted, and ANOVA followed by a Tukey’s post hoc test was used for multiple group comparisons (B-D, F, H) . The Kaplan–Meier method was used to estimate survival functions, and the log-rank test was used for group comparisons (I) . *p < 0.05; ns, not significant. CRPC, castration-resistant prostate cancer; PSMA, prostate-specific membrane antigen; Ab, antibody; PSA, prostate-specific antigen; IL-6, interleukin-6.

Article Snippet: Anti-PSMA Ab (10 mg/kg), IgG1 isotype (Med Chem Express, HY-P99001; 10 mg/kg), or PBS were administered intraperitoneally on days 10 and 18 after tumor inoculation.

Techniques: In Vivo, Injection, Control, Tumor Implantation, Membrane

Journal: Journal of Neuroinflammation

Article Title: CD22 modulation alleviates amyloid β-induced neuroinflammation

doi: 10.1186/s12974-025-03361-2

Figure Lengend Snippet: Suciraslimab suppresses Aβ-induced inflammation in microglia and human PBMC. A Effect of CD22 overexpression on Aβ-induced NFκB signaling in HEK293. Two-way ANOVA: CD22 expression, F (1,20) = 62.97, i < 0.0001; Aβ treatment, F (4,20) = 16.83, P < 0.0001. Tukey post hoc test, **** P < 0.0001. N = 3. B Effect of suciraslimab on Aβ-induced IL-1β release in MDMi. One-way ANOVA, F = 7.767, P = 0.004. Tuley post hoc test, * P < 0.05, ** P < 0.01. N = 6–7. C Immunofluorescent staining and quantitation of NLRP3 and ASC after Aβ and suciraslimab treatment in MDMi. NLRP3: One-way ANOVA, F = 10.09, P < 0.0001, Tukey post hoc test * P < 0.05, *** P < 0.001; ASC, One-way ANOVA, F = 19.10, P < 0.0001, Tukey post hoc test **** P < 0.0001. N = 6–15. D Effect of suciraslimab on Aβ-induced IL-1β release in human PBMC. One-way ANOVA, F = 6.833, P = 0.0052. Tukey post hoc test, ** P < 0.01, ns = not significant. N = 8. E Effect of suciraslimab on IFNγ + LPS-induced IL-23 and IL-12 release in human PBMC. IL-23: One-way ANOVA, F = 26.93, P = 0.0002. Tukey post hoc test, ** P < 0.01, *** P < 0.001; IL-12: One-way ANOVA, F = 10.21, P = 0.0008. Tukey post hoc test, * P < 0.05, *** P < 0.001. N = 4–8. F Effect of suciraslimab on TLR4 surface expression on monocyte upon IFNγ and LPS activation. Two-tailed paired Student’s t test, P = 0.0293, t = 3.322, df = 4. N = 5 G Effect of suciraslimab on α4 integrin surface expression on T cell of human PBMC. One-way ANOVA, F = 0.7059, P = 0.5131. N = 5. H Effect of suciraslimab on α4 integrin surface expression on B cell of human PBMC. One-way ANOVA, F = 66.02, P < 0.0001. Tukey’s post hoc test, * P < 0.05, **** P < 0.0001. N = 4–5. I Effect of suciraslimab on α4 integrin surface expression on T cell-depleted human PBMC. One-way ANOVA, F = 16.91, P = 0.0009. Tukey’s post hoc test, ** P < 0.01. N = 4. J Effect of suciraslimab on α4 integrin surface expression on monocyte-depleted human PBMC. One-way ANOVA, F = 8.565, P = 0.0083. Tukey’s post hoc test, IgG1 vs. αCD22 Ab, P = 0.1748. N = 4. All data are presented as mean ± SEM

Article Snippet: The cells were incubated overnight at 4 °C with anti Aβ antibody, clone 6E10 (BioLegend, 803014) and anti-CD22 antibody (SM03) or IgG1 isotype (Sino Biological, HG1K) for the control group.

Techniques: Over Expression, Expressing, Staining, Quantitation Assay, Activation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

doi: 10.3389/fimmu.2018.01848

Figure Lengend Snippet: Activation of 4-1BB signaling promotes the secretion of pro-fibrotic mediators by MH-S cells. MH-S cells treated with or without agonist 4-1BB mAb (10 µg/mL) or IgG (10 µg/mL) for 2 h prior to exposure to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) The levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E) Western blots analysis of IκBα and phospho-IκBα. (F) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (G–I) Real-time polymerase chain reaction analysis of MMP12, MMP9, and monocyte chemoattractant protein-1 mRNA expression ( n = 4). (J–L) ELISA analysis was used to quantify the secretion of IL-1β, IL-6 and tumor necrosis factor-α ( n = 4). The results were representative of three independent experiments. Results were graphed as the mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001).

Article Snippet: 100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

Techniques: Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

doi: 10.3389/fimmu.2018.01848

Figure Lengend Snippet: Blockade of 4-1BB signaling attenuates the secretion of pro-fibrotic mediators by MH-S cells. Transfected (Len-cont. and sh-4-1BB) MH-S cells were treated with or without NQDI 1 (10 µM), 4-1BBIg (10 µg/mL), or IgG1 (10 µg/mL) for 2 h, then exposed to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) Levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E,G) Western blots analysis of IκBα and phospho-IκBα. (F,H) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (I–K) The expressions of MMP12, MMP9, and monocyte chemoattractant protein-1 were detected by real-time polymerase chain reaction analysis ( n = 4). ELISA analysis of cytokines in the culture supernatants. (L) IL-1β, (M) IL-6, (N) tumor necrosis factor-α ( n = 4). Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant). The data were representative of three independent experiments.

Article Snippet: 100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

Techniques: Transfection, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

doi: 10.3389/fimmu.2018.01848

Figure Lengend Snippet: The secretion of pro-fibrotic mediators is reduced in the lungs from crystalline silica (CS)-injured mice, upon inhibition of 4-1BB signaling. C57BL/6 mice were administered a CS suspension or saline, respectively; 4-1BBIg or isotype control (IgG1) were injected intraperitoneally (i.p.; n = 3–4). (A–D) Quantification of MMP9 and MMP12 protein levels by western blot, which were normalized to those of β-actin in lungs. Shown as bar graph. (E–H) ELISA analysis of cytokines in lung tissues. (E) IL-1β, (F) IL-6, (G) tumor necrosis factor-α, (H) monocyte chemoattractant protein-1. Experiments were performed three times. C57BL/6 mice were administered a CS suspension or saline, respectively; NQDI 1 or isotype control were injected i.p. ( n = 10). (I) Immunohistochemical staining of paraffin-embedded lung tissue sections at 7 and 56 days showed CD68, MMP9, and MMP12 expression. Nuclei were stained by hematoxylin (blue). (J–L) Identification of MMP9 and MMP12 protein levels in mouse lung tissues at 7 and 56 days by western blot. The levels of MMP9 and MMP12 were normalized to those of β-actin. (M) Representative images for the immunohistochemical staining of collagen I in paraffin-embedded lung tissue sections 56 days after CS instillation. Nuclei were stained by hematoxylin (blue). (I,M) Scale bar, 50 µm. Experiments were performed three times. Data are shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant).

Article Snippet: 100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

Techniques: Inhibition, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing

Journal: PLoS ONE

Article Title: Frequency and role of NKp46 and NKG2A in hepatitis B virus infection

doi: 10.1371/journal.pone.0174103

Figure Lengend Snippet: Purified NK cells from HS were co-cultured with HepG2 (gray line/bar) or HepG2.2.15 (black line/bar). (A) Specific lysis of HepG2 or HepG2.2.15 and IFN-γ in the supernatant were assessed in indicating E/T ratio. ★ ; P<0.05 by unpaired Student’s t-test between HepG2 or HepG2.2.15. (B) NKp46-ligand expression in HepG2 or HepG2.2.15. (C) Specific lysis of HepG2 or HepG2.2.15 and IFN-γ in the supernatant were assessed when neutralizing antibody against NKp46 (Anti-NKp46 antibody) or isotype human IgG1 (Isotype IgG1) were added before co-culturing. *; P<0.05 by Kruskal-Wallis test.

Article Snippet: For the secondary antibody, these cells were stained with APC conjugated anti-human IgG1 antibody (97924; R&D systems) for 30 minutes and analyzed by flow cytometry.

Techniques: Purification, Cell Culture, Lysis, Expressing

Journal: PLoS ONE

Article Title: A Fully Human Inhibitory Monoclonal Antibody to the Wnt Receptor RYK

doi: 10.1371/journal.pone.0075447

Figure Lengend Snippet: (A) Flow cytometry using purified mouse anti-RYK MAbs 1B4, 1G8, 5E3 and 6G1 on 293-EBNA cells stably expressing pVITRO3-mcs (empty vector control; V) or hRYKFCT (RYK). All antibodies detected RYK in hRYKFCT-transfected but not vector-transfected cells. (B) Schematic of the mouse Ryk fusion proteins used in this study. EC, extracellular region; WD, WIF domain. (C) Western blot analysis of purified mouse Ryk fusion proteins using mouse anti-RYK MAbs 1B4 and 6G1. The pattern of binding was the same for both antibodies. The presence of all the fusion proteins was confirmed by stripping the membrane and reprobing with rabbit anti-Ryk EC polyclonal antibody. Molecular mass standards are shown at left in kDa. IB, immunoblot. (D) ELISA results using mouse anti-RYK MAbs 1B4 and 6G1 on an immobilized peptide library of the entire human RYK extracellular region. Peptides 3 to 37: RYK WIF domain; peptide 47: FLAG epitope (incubated with mouse anti-FLAG M2 MAb; positive control); well 48: empty (negative control). The MAbs were used at 2 µg/mL. All antibodies bound to the same epitope, in peptides 40−42. The location of the epitope is shown schematically (bottom). Epitopes for the 1G8 and 5E3 antibodies were identical (data not shown). OD, optical density.

Article Snippet: Cis -diols on a fully human anti-hapten IgG 1κ MAb (Eureka Therapeutics) were oxidized as above then biotinylated in 50 mM sodium phosphate, 150 mM NaCl, pH 7.0 at 23°C and exchanged into PBS for use as an isotype control (3.1 mol biotin/mol MAb) for IHC.

Techniques: Flow Cytometry, Purification, Stable Transfection, Expressing, Plasmid Preparation, Control, Transfection, Western Blot, Binding Assay, Stripping Membranes, Membrane, Enzyme-linked Immunosorbent Assay, FLAG-tag, Incubation, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: A Fully Human Inhibitory Monoclonal Antibody to the Wnt Receptor RYK

doi: 10.1371/journal.pone.0075447

Figure Lengend Snippet: (A) Co-IP of Wnt1.Myc5, Wnt3a.Myc5 and Wnt5a.Myc5 from lysates (200 µg) of transiently transfected HEK293T cells with 1 µg purified hRYKWD.Fc protein. Anti-FLAG IPs were immunoblotted (IB) with an anti-Myc antibody to detect co-precipitated Wnt. Molecular mass standards are shown at left in kDa. V, empty vector (pcDNA3)-transfected cells. (B) Anti-FLAG IPs were performed to pull down hRYK.Fc (250 ng), then immunoblotted (IB) as shown. Molecular mass standards are shown at left in kDa. IgG, human IgG control. (C) ELISA analysis of anti-RYK scFv proteins (5 µg/mL) on an immobilized peptide library of the entire human RYK extracellular region (peptides 1 to 46; peptide 47: FLAG epitope; well 48: empty). No linear epitopes were identified. OD, optical density.

Article Snippet: Cis -diols on a fully human anti-hapten IgG 1κ MAb (Eureka Therapeutics) were oxidized as above then biotinylated in 50 mM sodium phosphate, 150 mM NaCl, pH 7.0 at 23°C and exchanged into PBS for use as an isotype control (3.1 mol biotin/mol MAb) for IHC.

Techniques: Co-Immunoprecipitation Assay, Transfection, Purification, Plasmid Preparation, Control, Enzyme-linked Immunosorbent Assay, FLAG-tag

Journal: PLoS ONE

Article Title: A Fully Human Inhibitory Monoclonal Antibody to the Wnt Receptor RYK

doi: 10.1371/journal.pone.0075447

Figure Lengend Snippet: (A) RWD1 IPs from lysates of HEK293T cells transiently transfected with vectors encoding human RYK domain swap derivatives (lanes 1–5) were immunoblotted (IB) with anti-FLAG antibody. Lysate input (lanes 6–9) is shown at right. Molecular mass standards are shown at left in kDa. WIF1-WD, WIF domain from human WIF1; ROR2-CRD, CRD from human ROR2; H, heavy chain; L, light chain. (B) Anti-FLAG IPs were immunoblotted (IB) with an anti-Myc antibody to detect Wnt binding to hRYK.Fc. Molecular mass standards are shown at left in kDa. (C) ELISA analysis of immobilized RWD1 probed with hRYK.Fc. Increased hRYK.Fc binding to the antibody was observed with higher concentrations of either immobilized RWD1 (left panel) or soluble hRYK.Fc (right panel). Results represent the mean±standard deviation of two or three independent experiments. IgG, human IgG; OD, optical density. (D) HEK293T cells transiently transfected with the plasmids indicated (at right) were fixed as shown (above), paraffin-embedded and subjected to IHC using RWD1 biotinylated on N -glycan chains. Bar in panel (iv) represents 50 µm. NBF, neutral-buffered formalin; PFA, paraformaldehyde. (E) Example of IHC on a tumor from a formalin-fixed and paraffin-embedded human breast cancer tissue microarray stained with bRWD1 (upper panel) or human MAb isotype control (bIgG 1κ ; lower panel). Open arrow, RYK on a cancer cell; filled arrow, RYK on tumor stroma; arrowhead, RYK-positive cancer cell nucleus; b, biotinylated on N -glycan chains. Bar represents 50 µm.

Article Snippet: Cis -diols on a fully human anti-hapten IgG 1κ MAb (Eureka Therapeutics) were oxidized as above then biotinylated in 50 mM sodium phosphate, 150 mM NaCl, pH 7.0 at 23°C and exchanged into PBS for use as an isotype control (3.1 mol biotin/mol MAb) for IHC.

Techniques: Transfection, Binding Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Glycoproteomics, Microarray, Staining, Control