epha2 protein cf  (R&D Systems)

Journal: Theranostics

Article Title: EphA2-targeted alpha-particle theranostics for enhancing PDAC treatment

doi: 10.7150/thno.106948

Figure Lengend Snippet: CCLE and TCGA database of mRNA expression of EphA2. A) EphA2 expression in cell lines in CCLE. B) EphA2 mRNA expression in different types of tumors in TCGA. C) Comparison of mRNA expression of EphA2 in different normal tissues and corresponding tumors. D) Expression of different EPH receptors in pancreatic tumors. PAAD = pancreatic cancer; BLCA = Bladder cancer; READ = Rectal cancer; LUSC = Lung squamous cell carcinoma; THCA = Thyroid cancer; KIRP = Kidney papillary cell carcinoma; LUAD = Lung adenocarcinoma; KIRC = Kidney clear cell carcinoma; CHOL = Bile duct cancer; SKCM = melanoma; LIHC = Liver cancer; KICH = Kidney chromophobe; BRCA = Breast cancer; PRAD = Prostate cancer; DLBC = Large-B-cell lymphoma; LAML = Acute myeloid leukemia.

Article Snippet: His-tagged human EphA2 (R&D systems, catalog # 3035-A2, 56.9 kDa, 4.46 μM stock concentration) and mouse EphA2 (SinoBiological, cat.# 50586-M08H, 58 kDa, 8.6 μM stock concentration) were immobilized onto the CM5 chip.

Techniques: Expressing, Comparison

Journal: Theranostics

Article Title: EphA2-targeted alpha-particle theranostics for enhancing PDAC treatment

doi: 10.7150/thno.106948

Figure Lengend Snippet: Structure and in vitro characterization of AJ201. A) Structure of bicyclic peptide AJ201 having NOTA as bifunctional chelator for 68 Ga-labeling. In the structure, black color represents binding moiety, parakeet color represents the linker and red color represents the radiometal B) Surface plasmon resonance (SPR) analysis showing affinity of AJ201 for EphA2 using recombinant human (solid) and mouse (dashed) EphA2 proteins.

Article Snippet: His-tagged human EphA2 (R&D systems, catalog # 3035-A2, 56.9 kDa, 4.46 μM stock concentration) and mouse EphA2 (SinoBiological, cat.# 50586-M08H, 58 kDa, 8.6 μM stock concentration) were immobilized onto the CM5 chip.

Techniques: In Vitro, Labeling, Binding Assay, SPR Assay, Recombinant

Journal: Theranostics

Article Title: EphA2-targeted alpha-particle theranostics for enhancing PDAC treatment

doi: 10.7150/thno.106948

Figure Lengend Snippet: In vitro specificity of [ 68 Ga]AJ201 for EphA2 in human pancreatic cancer cells. A) Quantification of EphA2-mRNA using RT-PCR. B) Flow cytometry analysis of EphA2 receptor expression on cell surface. C) A representative plot of EphA2 receptor density in PDAC cells measured by quantibrite assay. D) [ 68 Ga]AJ201 binding (percent incubated activity, %IA) to different cells. Cells were incubated with 1 µCi [ 68 Ga]AJ201 at 4 °C for 1 h. [ 68 Ga]AJ201 uptake is EphA2 expression dependent, and co-incubation with 2 μM of non-radioactive AJ201 (0.2 nmol, blocking dose) significantly reduced radiotracer uptake confirming EphA2 specificity. E) In vitro uptake of [ 68 Ga]AJ201 over 60 min in Panc1 cells at 4 ˚C and 37 ˚C. F) Correlation of [ 68 Ga]AJ201 uptake with surface EphA2 receptor density. Data in panels A , D , E and F are presented as mean ± SD (n = 3-4). Significance was calculated using multiple unpaired t test in D ; ns, P ≥ 0.05; *, P ≤ 0.05; ***, P ≤ 0.001; ****, P ≤ 0.0001. Simple linear regression and Pearson coefficient were used in F . All in vitro experiments were performed three times and representatives are shown.

Article Snippet: His-tagged human EphA2 (R&D systems, catalog # 3035-A2, 56.9 kDa, 4.46 μM stock concentration) and mouse EphA2 (SinoBiological, cat.# 50586-M08H, 58 kDa, 8.6 μM stock concentration) were immobilized onto the CM5 chip.

Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry, Expressing, Binding Assay, Incubation, Activity Assay, Blocking Assay

Journal: Theranostics

Article Title: EphA2-targeted alpha-particle theranostics for enhancing PDAC treatment

doi: 10.7150/thno.106948

Figure Lengend Snippet: In vivo specificity of [ 68 Ga]AJ201 for EphA2 in NSG mice with PDAC tumor xenografts. A) Whole-body PET/CT images of different human PDAC xenografts at 60 min after the injection of radiotracer. Mice were injected with ~ 7.4 MBq (~200 µCi) [ 68 Ga]AJ201; K, kidney. Panc1 mouse image is reproduced from figure E. B) [ 68 Ga]AJ201 uptake quantification (%ID/g) in different PDAC tumors by ex vivo biodistribution at 60 min after injection. Left: %ID/g of different tumor xenografts, tumor-to-blood (middle) and tumor-to-muscle (right) ratios in each individual mice harboring respective xenografts. C) IHC staining for EphA2 expression in PDAC xenografts (digitally scanned at 40x).; data in figure B is shown as box and whisker plots (median ± IQR) showing all data points (n = 4-5).

Article Snippet: His-tagged human EphA2 (R&D systems, catalog # 3035-A2, 56.9 kDa, 4.46 μM stock concentration) and mouse EphA2 (SinoBiological, cat.# 50586-M08H, 58 kDa, 8.6 μM stock concentration) were immobilized onto the CM5 chip.

Techniques: In Vivo, Positron Emission Tomography-Computed Tomography, Injection, Ex Vivo, Immunohistochemistry, Expressing, Whisker Assay

Journal: Theranostics

Article Title: EphA2-targeted alpha-particle theranostics for enhancing PDAC treatment

doi: 10.7150/thno.106948

Figure Lengend Snippet: In vivo distribution of [ 68 Ga]AJ201 in Panc1-orthotopic tumor model. A) Coronal sections of the fused PET/MR images showing [ 68 Ga]AJ201 distribution. Primary tumor is indicated by red circle; K = Kidney, B = Bladder. B) Quantification of accumulated activity in tumor, muscle and pancreas of images shown in panel A (n = 4-5) C) tumor-to-muscle (T/M) and tumor-to-pancreas (T/P) ratios from the images shown in panel A (n = 4) D) EphA2 IHC of orthotopic Panc1 tumor and normal pancreas to validate the EphA2 expression. Panc1 orthotopic tumor model was generated by surgically implanting a 3-5 mm³ section of Panc1 tumor xenograft directly onto the pancreas. The implanted tumors were allowed to grow for 10 days, after which they were ready for imaging. At this stage, MRI measurements indicated that tumor sizes ranged from 10-50 mm 3 . data in figure B and C is shown as box and whisker plots (median ± IQR) showing all data points (n = 3-5).

Article Snippet: His-tagged human EphA2 (R&D systems, catalog # 3035-A2, 56.9 kDa, 4.46 μM stock concentration) and mouse EphA2 (SinoBiological, cat.# 50586-M08H, 58 kDa, 8.6 μM stock concentration) were immobilized onto the CM5 chip.

Techniques: In Vivo, Positron Emission Tomography-Magnetic Resonance Imaging, Activity Assay, Expressing, Generated, Imaging, Whisker Assay