Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 4. CML triggered by anti–EGFR-IgG3 negatively correlates with CD55 and CD59 expression levels. (A) Surface expression levels of CD46, CD55, and CD59 on analyzed cell lines were quantified by calibrated flow cytometry. Means 6 SEM of at least three independent experiments are presented. (B) Correlations between CD46, CD55, or CD59 and anti–EGFR-IgG3–mediated CDC were calculated for all four cell lines. CDC results at 2 mg/ml Ab concentration were taken from experiments presented in Fig. 2. (C) A431 cells were seeded into 10-cm plates and grown overnight. On the following day, cells were transfected with 50 nM control siRNA or with single siRNAs specific for CD46, CD55, CD59, or with a combination of all three mCRP-specific siRNAs for 72 h. Efficiency of siRNA-induced knockdown was analyzed by direct flow cytometry using fluorochrome-labeled, mCRP- specific Abs (CD46-Pacific blue, CD55-PE, CD59-FITC), or respective control Abs. (D–G) CDC against control siRNA or mCRP-specific, siRNA- transfected A431 cells was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human serum and anti–EGFR-IgG1 (upper panels), anti– EGFR-IgG3 (lower panels), as well as the respective control Abs at increasing concentrations. (H) Concentration-dependent binding of CD55-Ab (BRIC216, mouse IgG1) to A431 cells was analyzed by indirect immunofluorescence. Results from one representative experiment are presented. (I) CDC triggered by anti–EGFR-IgG1, anti–EGFR-IgG3, or respective control Abs (all at 66.67 nM) against A431 cells in the presence of saturating concentrations of CD55-Ab or a control Ab (both at 66.67 nM) was analyzed by 3-h [51Cr] release assays in the presence of 25% v/v human (Figure legend continues)

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Expressing, Cytometry, Concentration Assay, Transfection, Control, Knockdown, Labeling, Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 5. CD55 dampens anti–EGFR-IgG3–triggered CML and promotes C1q-dependent induction of AP amplification. BHK-EGFR+ #5 cells were transiently transfected with a control vector or a CD55 vector for 48 h. (A) Cell-surface expression of CD55 was analyzed by direct flow cytometry using PE-conjugated CD55-specific or control Abs. (B–D) The influence of CD55 overexpression on anti–EGFR-IgG3–mediated CDC was investigated by [51Cr] release assays either (B) in an Ab concentration–response curve, (C) in a time-dependent manner, or (D) in serum titration experiments. (E–H) The influence of the alternative complement pathway inhibitor CRIg (E and G), the presence of C1q in serum (F; at 66.67 nM Ab concentration; mean 6 SEM of triplicates), as well as of factor B (H; 13.33 nM Ab concentration, 12.5% v/v factor B–depleted serum, 200 mg/ml factor B), on anti–EGFR-IgG3–mediated CDC was analyzed using either (E and F) control vector–transfected or CD55 vector–transfected BHK-EGFR+ #5 cells or (G and H) DiFi cells (66.67 nM Ab concentration). (I) Deposition of factor Bb on control vector– or CD55 vector–transfected BHK-EGFR+ #5 cells was analyzed by flow cytometry. Relative deposition levels were calculated by equating RFI measured in the absence of Ab with 100%. Results are presented as mean 6 SEM of at least three independent experiments with different blood donors. *p # 0.05 anti–EGFR-IgG3 versus respective control Ab; (B–D, I) #p # 0.05 control vector versus CD55 vector; (E and G) #p # 0.05 without CRIg-Fc versus CRIg-Fc; (H) #p # 0.05 w/o factor B versus with factor B.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Cytometry, Over Expression, Concentration Assay, Titration

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Epidermal growth factor receptor targeting IgG3 triggers complement-mediated lysis of decay-accelerating factor expressing tumor cells through the alternative pathway amplification loop.

doi: 10.4049/jimmunol.1400329

Figure Lengend Snippet: FIGURE 6. Overview of complement activation by human anti–EGFR-IgG3 in the context of CD55 expression. On CD55-deficient target cells (left panel), anti–EGFR-IgG3 mediates strong C3b but low C4b deposition and induces assembly of classical and alternative C3 convertases, predominantly resulting in the induction of fast and efficient CDC via the classical pathway of complement activation. In contrast, on CD55-expressing target cells (right panel), CD55 mainly accelerates the decay of low amounts of classical C3 convertases, leading to amplification of the AP and finally to slow and inefficient CDC induction.

Article Snippet: To block complement regulatory activity of CD55, we used mouse anti-human CD55 (66.67 nM, BRIC216, mouse IgG1; Bio-Rad) blocking mAb in CDC experiments at saturating concentrations.

Techniques: Activation Assay, Expressing

Journal: iScience

Article Title: CRISPR-Cas9 screening reveals a distinct class of MHC-I binders with precise HLA-peptide recognition

doi: 10.1016/j.isci.2024.110120

Figure Lengend Snippet: Receptor-ligand CRISPR-Cas9 activation screen reveals that CD55 interacts with HLA-C∗07:01-VRIG tetramers (A) Schematic of the receptor ligand CRISPR-Cas9 activation screen. K562 cells transduced with a genome-wide activation library were stained with a pool of three HLA tetramers (HLA-A∗02:01-NLVP, HLA-B∗07:02-TPRV, and HLA-C∗07:01-VRIG), and enriched gRNAs in stained cells were identified using NGS. (B) SigmaFC scores of genes from two replicate screens. SigmaFC scores were calculated using PinAplPy, and top hits are annotated. (C) K562 cells stably expressing dCas9 and transduced with a gRNA upregulating CD55 or a control guide were stained with the HLA-A, -B, -C, or tetramers as in (A) or with HLA-E∗01:01-VMAP tetramers and analyzed by flow cytometry. (D) In vitro co-immunoprecipitation of recombinant CD55-Fc with HLA-A∗02:01-NLVP, HLA-B∗07:02-TPRV, HLA-C∗07:01-VRIG, or HLA-E∗01:01-VMAP tetramers. (E) Three different cell lines (HeLa, PC-3M, or SiHa) that express CD55 endogenously were stained for CD55 (top) or with HLA-C∗07:01-VRIG tetramers (bottom) and analyzed by flow cytometry. (F) HeLa wild-type or HeLa CD55 KO cells were stained with αCD55 or HLA-C∗07:01-VRIG tetramers and analyzed by flow cytometry. All data except (B) represent at least three independent experiments. CRISPRa, CRISPR activation screen; TMs, tetramers; WT, wild-type; KO, knockout. Related to Figure S1 and Table S1 .

Article Snippet: Antibodies used in this study were anti-heparan sulfate chains (AMSBIO, F58-10E4), FITC anti-human CD55 (Biolegend, 311306), anti-human CD55 BRIC110 (ARP, 08-9402-2, targets SCR2 of CD55), anti-human CD55 BRIC216 (Biorad, MCA914T, targets SCR3 of CD55), anti-human CD55 MAB2009 (R&D systems, MAB2009-SP, targets SCR1 of CD55), anti-SDC2 APC (R&D systems, FAB2965A), anti-SDC4 APC (R&D systems, FAB29181A), anti-CD55 APC (Biolegend, #311311), or goat-anti-mouse APC (Biolegend, #405308).

Techniques: CRISPR, Activation Assay, Transduction, Genome Wide, Staining, Stable Transfection, Expressing, Control, Flow Cytometry, In Vitro, Immunoprecipitation, Recombinant, Knock-Out

Journal: iScience

Article Title: CRISPR-Cas9 screening reveals a distinct class of MHC-I binders with precise HLA-peptide recognition

doi: 10.1016/j.isci.2024.110120

Figure Lengend Snippet: Interaction of CD55 with HLA-C∗07:01-VRIG tetramers is allotype and peptide specific (A) HEK293T cells were transfected with a plasmid containing GFP and a truncation mutant of CD55 and analyzed by flow cytometry. GFP+ positive cells were analyzed for staining with HLA-C∗07:01-VRIG. Each mutant removes an additional SCR domain from CD55. Data are represented as mean ± SD. (B) HeLa cells were stained with HLA-C∗07:01-VRIG tetramers after pre-incubation with CD55 blocking antibodies targeting different SCR domains on CD55 and analyzed by flow cytometry. (C) HeLa cells were stained with either HLA-C∗07:01 or HLA-C∗07:02 tetramers loaded with the VRIG peptide and analyzed by flow cytometry. (D) HeLa cells were stained with HLA-C∗07:01 tetramers loaded with different alanine mutants of the VRIGHLYIL peptide and analyzed by flow cytometry. (E) CD55-Fc was immobilized on a Prot-G chip for SPR data using HLA-C∗07:01-VRIG tetramers as analyte to determine interaction on and off rates and K D . Response units were measured with increasing concentrations of HLA-C∗07:01-VRIG tetramers. All data represent at least three independent experiments, except (E), which represents a biological duplicate. FL, full length. Related to Figure S2 , Tables S2 and .

Article Snippet: Antibodies used in this study were anti-heparan sulfate chains (AMSBIO, F58-10E4), FITC anti-human CD55 (Biolegend, 311306), anti-human CD55 BRIC110 (ARP, 08-9402-2, targets SCR2 of CD55), anti-human CD55 BRIC216 (Biorad, MCA914T, targets SCR3 of CD55), anti-human CD55 MAB2009 (R&D systems, MAB2009-SP, targets SCR1 of CD55), anti-SDC2 APC (R&D systems, FAB2965A), anti-SDC4 APC (R&D systems, FAB29181A), anti-CD55 APC (Biolegend, #311311), or goat-anti-mouse APC (Biolegend, #405308).

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Flow Cytometry, Staining, Incubation, Blocking Assay

Journal: iScience

Article Title: CRISPR-Cas9 screening reveals a distinct class of MHC-I binders with precise HLA-peptide recognition

doi: 10.1016/j.isci.2024.110120

Figure Lengend Snippet:

Article Snippet: Antibodies used in this study were anti-heparan sulfate chains (AMSBIO, F58-10E4), FITC anti-human CD55 (Biolegend, 311306), anti-human CD55 BRIC110 (ARP, 08-9402-2, targets SCR2 of CD55), anti-human CD55 BRIC216 (Biorad, MCA914T, targets SCR3 of CD55), anti-human CD55 MAB2009 (R&D systems, MAB2009-SP, targets SCR1 of CD55), anti-SDC2 APC (R&D systems, FAB2965A), anti-SDC4 APC (R&D systems, FAB29181A), anti-CD55 APC (Biolegend, #311311), or goat-anti-mouse APC (Biolegend, #405308).

Techniques: Virus, Recombinant, Blocking Assay, Genome Wide, Activation Assay, CRISPR, Knock-Out, Mutagenesis, Plasmid Preparation, Software, Imaging

Journal: The FASEB Journal

Article Title: MAP‐2:CD55 chimeric construct effectively modulates complement activation

doi: 10.1096/fj.202300571r

Figure Lengend Snippet: FIGURE 1 Expression of the novel construct MAP-2:CD551-4. (A) Graphical representation of MAP-2 and MAP-2:CD551-4. (B–D) Western immunoblotting of the purified protein MAP-2:CD551-4 probed with mAb anti-MASP-2/Map19 (B), mAb anti-CD55 (C), and mAb anti-FLAG tag (D). rMAP-2 and rCD551-4 produced in-house were used as controls. (E) Direct protein stain of the same purified proteins.

Article Snippet: Membranes were blocked with 5% (w/v) skim milk (Sigma- Aldrich) for 30 min and then incubated with 0.5 μg/mL rat monoclonal antibody (mAb) anti- MASP- 2/ Map19, clone 6G12 (Hycult, Uden, The Netherlands), 1 μg/mL mouse mAb anti- CD55, clone 278803 (MAB2009; R&D Systems, Minneapolis, MN, USA), or 2 μg/mL mouse mAb anti- FLAG- tag (clone 18, produced in- house).

Techniques: Expressing, Construct, Western Blot, Purification, FLAG-tag, Produced, Staining

Journal: The FASEB Journal

Article Title: MAP‐2:CD55 chimeric construct effectively modulates complement activation

doi: 10.1096/fj.202300571r

Figure Lengend Snippet: FIGURE 2 Structural characterization of MAP-2:CD551-4. (A) MAP-2:CD551-4 elution profile from size exclusion chromatography exposed to 2 mM of calcium, 10 mM EDTA, or 10 mM EGTA. Vertical lines represent the molecular weight of the molecular marker, represented in kDa. (B–D) Western immunoblotting of the elution fractions 3–16 of MAP-2 from SEC exposed to 2.5 mM of calcium (B), 10 mM EDTA (C), or 10 mM EGTA (D). MAb anti MASP-2/Map19 clone 6G12 was used to detect MAP-2:CD551-4. Representative blots out of 3 independent replicates are shown. (E and F) Thermal stability of MAP-2, CD55, and MAP-2:CD551-4 exposed to 2 mM calcium or 10 mM EGTA shown as the ratio of the intrinsic fluorescence at 350 and 330 nm (E) and the right the first derivative of the ratio (F). Thermal stability was performed in triplicates.

Article Snippet: Membranes were blocked with 5% (w/v) skim milk (Sigma- Aldrich) for 30 min and then incubated with 0.5 μg/mL rat monoclonal antibody (mAb) anti- MASP- 2/ Map19, clone 6G12 (Hycult, Uden, The Netherlands), 1 μg/mL mouse mAb anti- CD55, clone 278803 (MAB2009; R&D Systems, Minneapolis, MN, USA), or 2 μg/mL mouse mAb anti- FLAG- tag (clone 18, produced in- house).

Techniques: Size-exclusion Chromatography, Molecular Weight, Marker, Western Blot, Fluorescence

Journal: Pathogens

Article Title: Relationship Between Cell Surface Viral Glycoprotein Expression and Resistance of Parainfluenza Virus Persistently Infected Cells to Complement-Mediated Lysis

doi: 10.3390/pathogens14080815

Figure Lengend Snippet: C′ regulators are not significantly upregulated in H1975 and HEp2 PI cells. ( A , B ) RNA samples from H1975 ( A ) and HEp2 ( B ) cells were collected as described in the legend to . Samples were analyzed via RT-qPCR to determine gene expression of three C′ regulators: CD46, CD55, and CFH. Values were normalized to expression of β-actin. C′ inhibitor expression in mock-infected cell cultures was set equal to 1 and expression in AI and PI cel cultures is expressed as fold change relative to mock. ( C ) Mock-infected, AI, and PI HEp2 cells were stained with an anti-CD55 antibody and processed via flow cytometry. MFI; Mean Fluorescence Intensity. ns stands for not significant, **** indicates p -value < 0.0001.

Article Snippet: CD55 surface expression was monitored using an AF405 conjugated anti-human CD55 antibody (R&D Systems, FAB20091V).

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, Infection, Staining, Flow Cytometry, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Complement-Mediated Two-Step NETosis: Serum-Induced Complement Activation and Calcium Influx Generate NADPH Oxidase-Dependent NETs in Serum-Free Conditions

doi: 10.3390/ijms25179625

Figure Lengend Snippet: Complement activation increases complement regulators on neutrophils. Neutrophils were precoated with anti-CD59 monoclonal antibodies and then exposed to normal human serum (NHS) to activate the complement cascade (complement stimulated). The group not treated with anti-CD59 monoclonal antibodies served as the unstimulated control (serum). The cells were then fixed, immunostained, and analyzed by flow cytometry. The expression level of all three complement regulators (CD46, CD55, and CD59) was significantly increased on complement-activated neutrophils compared to the unstimulated controls. ( A ) Representative flow cytometry tracings of an experiment; ( B ) mean fluorescence intensity (MFI) for each marker from all experiments. n = 3 biological replicates. * p < 0.05, compared to their controls, based on the paired t-test. Data are presented as means ± SD.

Article Snippet: To specifically deactivate the complement activity of serum, while leaving other serum proteins functional, serum was heated for 56 degrees Celsius for 30 min. Primary antibodies C3b (1:500; Mouse, ThermoFisher Invitrogen, Carlsbad, CA, USA; Cat# MA1-70054), C5b-9 (1:250, Rabbit, Abcam, Toronto, ON, Canada; Cat# Ab55811), CD46 (1:200, Rabbit, Santa Cruz Biotech, Dallas, TX, USA; Cat# SC 9098), CD55 (1:250, Goat, R&D Systems, 614 McKinley Place NE, Minneapolis, MN, USA; Cat# AF2009), CD59 (1:200, Rabbit ThermoFisher Invitrogen, Carlsbad, CA, USA; Cat# PA5-34513), and PE CD11b (BioLegend, Markham, ON, Canada; Cat# 301306) and were incubated for 30 min on dark ice and then rinsed twice with spins at 400 g for 10 min.

Techniques: Activation Assay, Bioprocessing, Control, Flow Cytometry, Expressing, Fluorescence, Marker

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: CD55 expression in human lung cancer tissue.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: CD55 expression in human lung cancer tissue. ( a ) Immunohistochemical analysis of CD55 in lung tissue. i, normal lung; ii, lung adenosquamous carcinoma; iii, lymph node metastatic carcinoma from lung adenocarcinoma; iv, lung mucoepidermoid carcinoma; v, lung squamous cell; vi, lung adenocarcinoma; vii, lung large cell carcinoma; viii, lung bronchioloalveolar carcinoma. Scale bars = 100 μm. Insets are magnified images demonstrating CD55 expression on the cell membrane and in the cytoplasm. ( b ) Immunohistochemical analysis of CD55 expression in normal organs. i, spleen; ii, lung; iii, skeletal muscle; iv, kidney; v, rectum; vi, colon; vii, stomach; viii, liver. Scale bars = 100 μm.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Expressing, Immunohistochemical staining, Membrane

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Development of chimeric CD55-specific monoclonal antibodies. ( a ) Phage ELISA results for individual clones tested against recombinant human CD55. A total of 384 clones were tested for binding to recombinant human CD55 (Supplementary Fig. ). The data for 36 clones are shown. ( b ) Reactivity of three anti-CD55 IgGs to recombinant human CD55 analyzed by antigen-binding ELISA. ( c ) SDS-PAGE analysis of the Ab1 anti-CD55 IgG antibody. ( d ) Representative flow cytometry analysis of H460 and H69 cells stained with the Ab1 anti-CD55 antibody.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Bioprocessing, Enzyme-linked Immunosorbent Assay, Clone Assay, Recombinant, Binding Assay, SDS Page, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Characterization of 177 Lu-DTPA-anti-CD55 in vitro . ( a ) Representative results of three independent Lindmo assays to examine binding of 177 Lu-anti-CD55 to H460 cells. ( b ) Representative results of three independent saturation binding assays of 177 Lu-anti-CD55 in H460 cells. ( c ) Blocking assays in H460, H358, H69, and WI-38 cells (n = 3; **P < 0.01; Student’s t test). The results are presented as the mean ± standard error of the mean (SEM; error bars).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vitro, Binding Assay, Blocking Assay

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Biodistribution analysis of 177 Lu-anti-CD55 in vivo . ( a ) The biodistribution of 177 Lu-anti-CD55 in tissue from pleural metastatic mice (n = 3–6 per condition). The results are presented as the mean ± SD (error bars). ( b ) The biodistribution of 177 Lu-anti-CD55 in normal lung and lung tumor tissue from pleural metastatic mice (n = 3–6 per condition; **P < 0.01; Student’s t test). The results are presented as the mean ± SD (error bars). ( c ) Excretion and residual radioactivity after injection of 177 Lu-anti-CD55 into mice. ( d ) Micro-SPECT/CT images of pleural metastatic or control mice treated with 177 Lu-anti-CD55. The amount of radioactivity is calculated in MBq/cc.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vivo, Radioactivity, Injection, Micro-SPECT, Control

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Biodistribution of the 177 Lu-anti-CD55 antibody in a pleural metastatic mouse model.

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques:

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Effects of the 177 Lu-anti-CD55 antibody on invasion and migration in lung cancer cells. ( a ) The invasive front of squamous cell carcinoma in the lung (arrows). Scale bars = 50 μm. ( b ) Invasion and migration of H460 cells treated with IgG, anti-CD55, or 177 Lu-anti-CD55. ( c ) Invasion and migration were quantified by counting cells in ten randomly selected regions (***P < 0.001; Student’s t test). The results are presented as the means ± SEM (error bars).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Migration

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Therapeutic efficacy of the 177 Lu-anti-CD55 antibody in vitro and in vivo . ( a ) Assays of H460 lung cancer cell viability after treatment with IgG, 177 Lu-IgG, anti-CD55, or 177 Lu-anti-CD55. α-CD55 (μg/ml) indicates the concentration of all the antibodies, IgG, 177 Lu-IgG, α-CD55, and 177 Lu-α-CD55. 177 Lu-α-CD55 (MBq) indicates the corresponding amounts of radioactivity of 177 Lu-α-CD55 for the indicated concentration (n = 3; **P < 0.01; Student’s t test). The results are presented as the mean ± SEM (error bars). ( b ) Assays of H358 cell viability after treatment with IgG, 177 Lu-IgG, anti-CD55, or 177 Lu-anti-CD55 (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( c ) Effects of 177 Lu-anti-CD55 on the survival of H460 bearing-early pleural metastatic mice (n = 10 for each group; ***P < 0.001; Log-rank (Mantel-Cox) test). ( d ) Effects of 177 Lu-anti-CD55 on the survival of pleural metastatic mice (n = 10 for each group; **P < 0.01; Log-rank (Mantel-Cox) test).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: Drug discovery, In Vitro, In Vivo, Concentration Assay, Radioactivity

Journal: Scientific Reports

Article Title: Development of a radionuclide-labeled monoclonal anti-CD55 antibody with theranostic potential in pleural metastatic lung cancer

doi: 10.1038/s41598-018-27355-8

Figure Lengend Snippet: Combinatorial effects of 177 Lu-anti-CD55 and cisplatin in vitro and in vivo . ( a ) Cell viability assays of H460 lung cancer cells treated with 0.1 MBq of 177 Lu-anti-CD55 in the presence or absence of 3 μM cisplatin (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( b ) Cell viability assays of H358 lung cancer cells treated with 0.1 MBq of 177 Lu-anti-CD55 in the presence or absence of 3 μM cisplatin (n = 3; *P < 0.05; Student’s t test). The results are presented as the mean ± SEM (error bars). ( c ) Effects of 177 Lu-anti-CD55 in the presence or absence of cisplatin on the survival of early pleural metastatic mice (n = 10 for each group; ***P < 0.001; Log-rank (Mantel-Cox) test). ( d ) Analysis of the survival of pleural metastatic mice after treatment with 177 Lu-anti-CD55 in the presence or absence of cisplatin (n = 10 for each group).

Article Snippet: Briefly, 5 × 10 6 magnetic beads (Dynabeads M-270 epoxy; Invitrogen) were coated with 2.5 μg of recombinant human CD55 (2009-CD/CF; R&D Systems) for each round of bio-panning (four rounds).

Techniques: In Vitro, In Vivo