Journal: Frontiers in Oncology

Article Title: Establishment, characterization, and drug screening of low-passage patient individual non-small cell lung cancer in vitro models including the rare pleomorphic subentity

doi: 10.3389/fonc.2023.1089681

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: LYPD3 , APC , Sino Biological Europe GmbH, Düsseldorf, Germany , 11836-H08H.

Techniques: Cytometry

Journal: Frontiers in Oncology

Article Title: Establishment, characterization, and drug screening of low-passage patient individual non-small cell lung cancer in vitro models including the rare pleomorphic subentity

doi: 10.3389/fonc.2023.1089681

Figure Lengend Snippet: Flow cytometry. Histogram overlays of unstained controls (dotted lines for all three cell lines) and measurements for HROLu55 (green), HROLu22 (blue), and HROBML01 (red) for the epitopes CD326, PD-L1, EGFR, CD26, LYPD3, DSG3, CCD59, CD27, and CD90 are shown.

Article Snippet: LYPD3 , APC , Sino Biological Europe GmbH, Düsseldorf, Germany , 11836-H08H.

Techniques: Flow Cytometry

Journal: Frontiers in Drug Discovery

Article Title: Targeting a cancer-specific LYPD3 glycoform for tumor therapy

doi: 10.3389/fddsv.2023.1298916

Figure Lengend Snippet: FIGURE 1 Glycosylation-dependent recognition of LYPD3 by GT-002. (A) LYPD3 proteins carrying truncated O-glycans (w/O-glyc) or no O-glycans (w/o O-glyc) were coated onto ELISA plates and tested for binding of 250 ng/mL GT-002 and Lupartumab, both expressed with human IgG1 backbones. (B) Dose-response curves (exemplary experiment measured in duplicates) of GT-002 (black circles) and Lupartumab (white squares) were determined by ELISA on LYPD3 carrying truncated O-glycans. (C) Flow cytometric analysis of cells overexpressing LYPD3 carrying truncated O-glycans (LYPD3-F9), no O-glycans (LYPD3-HEK-OglycKO) or low endogenous LYPD3 levels (F9) using 10 μg/mL GT-002. Lupartumab (1 μg/mL) and aTF mAb (0.1 μg/mL) served as expression controls. Signal/noise ratios were calculated as quotient of MFI (mean fluorescence intensity) values of mAb and appropriate isotype control staining. Error bars indicate standard deviation (SD) of duplicates.

Article Snippet: Protein-specific anti-LYPD3 polyclonal antibodies HPA041529 (Atlas Antibodies) or AF5428 (R&D Systems) served as positive control.

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

Journal: Frontiers in Drug Discovery

Article Title: Targeting a cancer-specific LYPD3 glycoform for tumor therapy

doi: 10.3389/fddsv.2023.1298916

Figure Lengend Snippet: FIGURE 2 Fine-specificity of GT-002 in ELISA. (A) O-glycan profiling of LYPD3 glycoforms used for binding studies. Monosaccharide symbols follow the CFG recommended symbol nomenclature (Varki et al., 2015). Open circles: unspecified hexose; peaks marked with an asterisk are methodically-derived peeling products. (B) Recognition of indicated LYPD3 glycoforms by GT-002 (12.5 ng/mL) was analyzed by ELISA. LYPD3 purified from different cell lines was treated with neuraminidase (de-sialyl.) or with galactosidase (de-gal.), where indicated. Lupartumab (10 ng/mL), aTF mAb (1 μg/mL) and aTn mAb (5 μg/mL) served as controls. (C) Binding of GT-002 (250 ng/mL) to irrelevant TF-carrying glycoproteins produced in F9 cells was tested by ELISA. Lupartumab (250 ng/mL) and aTF mAb (1 μg/mL) served as controls. Error bars indicate standard deviation (SD) of duplicates.

Article Snippet: Protein-specific anti-LYPD3 polyclonal antibodies HPA041529 (Atlas Antibodies) or AF5428 (R&D Systems) served as positive control.

Techniques: Enzyme-linked Immunosorbent Assay, Glycoproteomics, Binding Assay, Derivative Assay, Produced, Standard Deviation

Journal: Frontiers in Drug Discovery

Article Title: Targeting a cancer-specific LYPD3 glycoform for tumor therapy

doi: 10.3389/fddsv.2023.1298916

Figure Lengend Snippet: FIGURE 3 Binding of GT-002 to tumor cells. (A) LYPD3-positive (CaOV-3 and ZR-75-1) and -negative (MDA-MB-231) human tumor cell lines were treated with neuraminidase (w/neu) or not treated (w/o neu) and stained with 10 μg/mL GT-002. Lupartumab (1 μg/mL) and aTF (10 μg/mL) served as expression controls. Signal to noise ratios were calculated as the quotient of MFI values of mAb and appropriate isotype control stainings. (B) Immunohistochemical staining of a human tumor CDX section derived from the CaOV-3 cell line with 10 μg/mL GT-002. Scale bars indicate 50 µm. Error bars indicate SD of duplicates.

Article Snippet: Protein-specific anti-LYPD3 polyclonal antibodies HPA041529 (Atlas Antibodies) or AF5428 (R&D Systems) served as positive control.

Techniques: Binding Assay, Staining, Expressing, Control, Immunohistochemical staining, Derivative Assay

Journal: Frontiers in Drug Discovery

Article Title: Targeting a cancer-specific LYPD3 glycoform for tumor therapy

doi: 10.3389/fddsv.2023.1298916

Figure Lengend Snippet: FIGURE 6 Internalization of GT-002 into tumor cell line. GT-002 (black circles), Lupartumab (white squares) or an isotype control (grey circle) where labeled with anti-human Fabfluor-pH antibody labeling dye and then added to LYPD3-F9 cells (A) or F9 cells (B). Internalization, measured as red fluorescence induced by low acidic pH after antibody uptake, was monitored for 24 h. Percentage of lysosomal routing was expressed as the Red Area (Red positive cells)/Phase Area (Total cell confluency). Error bars indicate SD of duplicates.

Article Snippet: Protein-specific anti-LYPD3 polyclonal antibodies HPA041529 (Atlas Antibodies) or AF5428 (R&D Systems) served as positive control.

Techniques: Control, Labeling, Antibody Labeling

Journal: Antibodies

Article Title: A High-Affinity Monoclonal Antibody Against the Pancreatic Ductal Adenocarcinoma Target, Anterior Gradient-2 (AGR2/PDIA17)

doi: 10.3390/antib13040101

Figure Lengend Snippet: High-affinity anti-AGR2 MAbs inhibit AGR2-mediated cell adhesion, migration, and binding to LYPD3. ( A ) ELISA plates were coated with BSA or AGR2, and PANC-1 cells were allowed to adhere for 1 h in the absence (“0”) or presence of the indicated anti-AGR2 MAbs. Non-adherent cells were washed away, and adherent cells were quantified using the Cell Titer-Glo assay system. Cell adhesion data are shown. ( B ) KPC cells were used in migration assays using a Boyden chamber design with 8 μm porous membrane. (Left) KPC cells were added to the top chamber in the presence or absence of rAGR2 (500 nM) and allowed to migrate toward a chemoattractant gradient composed of KPC-conditioned media (CM) for 6 h. Quantification of migrated cells is shown. (Right) KPC or KPC cells with AGR2 overexpression were added to the top chamber and allowed to migrate toward a chemoattractant gradient composed of KPC-conditioned media (CM) for 6 h. Quantification of migrated cells is shown. Statistical significance was determined using a Student’s t -test (n = 3). ( C ) LYPD3 binding to AGR2-coated or BSA-coated (control) plates was analyzed. ELISA data are shown. ( D ) LYPD3 binding to AGR2-coated plates was measured in the presence of the indicated concentrations of the affinity-mature anti-AGR2 MAb S31R/A53Y. ELISA data are shown.

Article Snippet: Subcloning: Genes for AGR2, LYPD3, scFvs, and monoclonal antibodies were codon optimized for either E. coli or human cells by GenScript Biotech (Piscataway, NJ, USA.

Techniques: Migration, Binding Assay, Enzyme-linked Immunosorbent Assay, Glo Assay, Membrane, Over Expression, Control

Journal: Frontiers in Genetics

Article Title: LYPD3 , a New Biomarker and Therapeutic Target for Acute Myelogenous Leukemia

doi: 10.3389/fgene.2022.795820

Figure Lengend Snippet: (A) Expression of LYPD3 in cancer lines (the expression of LYPD3 is the highest in HL-60 cells); (B) LYPD3 gene knockdown mediated by siRNA-induced apoptosis in AML cells (HL-60 cells); (C) LYPD3 gene knockdown mediated by siRNA-suppressed proliferation in AML cells (HL-60 cells); (D) significantly enriched pathways in AML samples with high LYPD3 expression; (E) significantly enriched pathways (the P53 signaling pathways); (F) significantly enriched pathways (PI3K_AKT signaling pathway); (G) relationship between LYPD3 and the molecules Akt and P53 (the expression of the LYPD3 gene knockdown-mediated SiRNA group was obviously increased in p53 and PI3K_AKT signaling).

Article Snippet: We obtained LYPD3 siRNA and scrambled negative control siRNA for transfection from Shanghai GenePharma (China).

Techniques: Expressing