biotinylated recombinant human psma  (R&D Systems)

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: Schematic diagram to show PCa‐active targeting and combined anti‐tumor effects of P‐MMCNPs@DM1 after intravenous injection. P‐MMCNPs have a core‐shell structure, with a macrophage membrane that has TM‐expressed gy1‐enhanced green fluorescent protein (EGFP) fusion protein as the shell and FeAuNPs as the core. Then, the cytotoxic drug DM1 was loaded to obtain the P‐MMCNPs@DM1. When P‐MMCNPs@DM1 was injected into tumor‐bearing mice, the TM‐expressed gy1 could specifically recognize, bind with, and internalize into PSMA + PCa cells, in which DM1 was released from the lysosome to induce cytotoxicity. Together with PTT treatment, it could result in a combined anti‐tumor effect. In addition, the macrophage membranes of P‐MMCNPs also neutralize pro‐tumor cytokines and block the M2 macrophage‐mediated tumor‐promoting cascades.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: Injection, Membrane, Blocking Assay

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: Preparation and characterization of P‐MMCNPs. (A) Schematic diagram to show the design and fabrication of P‐MMCNPs. Anti‐PSMA scFv (gy1)‐EGFP fusion protein was stably TM‐expressed in RAW264.7 cells, and the cell membrane was extracted and assembled with FeAuNPs to obtain P‐MMCNPs. MM represents Raw264.7 gy‐1‐EGFP macrophage membrane, (B) Flow cytometry to show the expression of gy1‐EGFP fusion protein on Raw264.7 gy1‐EGFP cells, (C) Cellular ELISA to analyze the binding affinity of Raw264.7 gy1‐EGFP to PSMA. Data were analyzed using one‐way ANOVA, (D) TEM and SEM images of FeAuNPs, MM, and P‐MMCNPs. Pseudo‐color processing was employed to highlight the structural features. Scale bars, 500 nm, 100 nm, and 50 nm, and (E) Hydrodynamic diameter and zeta potential of FeAuNPs, MM, and P‐MMCNPs. Data were shown as mean ± SD from three independent experiments. **** P < 0.0001; Abbreviation : NS, no significant.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: Stable Transfection, Membrane, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay, Zeta Potential Analyzer

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs specifically bind with and internalize into PSMA + PCa cells. (A) Flow cytometry to examine the specific binding of P‐MMCNPs to PSMA + PCa cells, (B) Immunofluorescent staining to show the co‐localization of P‐MMCNPs and lysosomes in PSMA + PCa cells. Lysosome was stained with Lyso‐Tracker (red). Scale bar, 10 µm, and (C) Quantitative analysis of the fluorescent intensity ratio of Lyso‐Tracker to EGFP in PSMA + PCa cells. Data were analyzed using the student t ‐test (two‐tailed). Data were shown as mean ± SD from three independent experiments. *** P < 0.001.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: Flow Cytometry, Binding Assay, Staining, Two Tailed Test

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs specifically target PSMA + primary PCa in vivo. (A) Fluorescence imaging and quantification of fluorescence intensity to show the specific localization of P‐MMCNPs in subcutaneous PC3 PSMA+ tumor‐bearing mice. ( n = 3). Bioluminescence imaging (BLI) displayed the tumor site. Data were analyzed using repeated‐measures ANOVA, (B) Fluorescence imaging and quantification of fluorescence intensity to show the specific localization of P‐MMCNPs in isolated PC3 PSMA+ tumors. ( n = 3). Data were analyzed using the student t ‐test (two‐tailed), (C) CT imaging and quantitative analysis to show the specific localization of P‐MMCNPs in isolated PC3 PSMA+ tumors. ( n = 4). Untreated tumor‐bearing mice were used as controls. Data were analyzed using the student t ‐test (two‐tailed), (D) Dual‐modality MRI and quantitative analysis to show the specific localization of P‐MMCNPs in subcutaneous PC3 PSMA+ tumor‐bearing mice. ( n = 4). The white arrow indicates the tumor site. Data were analyzed using the student t ‐test (two‐tailed), and (E) The half‐life of P‐MMCNPs@DM1 in circulation after intravenous injection. Data were analyzed using repeated‐measures ANOVA. Data were shown as mean ± SD. **** P < 0.0001, *** P < 0.05, ** P < 0.01, and * P < 0.001; Abbreviation : NS, no significant.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: In Vivo, Fluorescence, Imaging, Isolation, Two Tailed Test, Injection

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs@DM1 alone or combined with PTT show potent cytotoxicity to PSMA + PCa cells in vitro. (A) CCK‐8 assay to show the specific cytotoxicity of P‐MMCNPs@DM1 on PSMA + PCa cells, (B) CCK‐8 assay to show the specific cytotoxicity of P‐MMCNPs@DM1 on PSMA + PCa cells compared with DM1. Untreated PCa cells were used as a control. Data were analyzed using the student t ‐test (two‐tailed), (C) Immunofluorescent staining of α‐tubulin in PC3 PSMA+ cells after incubation with P‐MMCNPs@DM1. Untreated PC3 PSMA+ cells were used as a control. The yellow rectangle indicates mitotic cells or damaged tubulin. Scale bars: 10 µm, (D) Quantification of α‐tubulin fluorescence intensity in (C). Data were analyzed using the student t ‐test (two‐tailed), (E) Quantification of mitotic cells in (C). Data were analyzed using the student t ‐test (two‐tailed), (F) CCK‐8 assay to show the photothermal cytotoxicity of P‐MMCNPs on PSMA + PCa cells after laser irradiation, and (G) CCK‐8 assay to show the DM1 and PTT combined anti‐tumor effects of P‐MMCNPs@DM1 on PSMA + PCa cells after laser irradiation. The laser intensity was 0.5 W · cm −2 and the irradiation time was 5 min. Data were analyzed using the student t ‐test (two‐tailed). Data were shown as mean ± SD from three independent experiments. **** P < 0.0001 and ** P < 0.01; Abbreviations : ND, not detected; NS, no significant.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: In Vitro, CCK-8 Assay, Control, Two Tailed Test, Staining, Incubation, Fluorescence, Irradiation

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs@DM1 in combination with PTT exerts a combined anti‐tumor effect on PSMA + tumors in vivo. (A) Schematic illustration to show the design of animal experiments in the subcutaneous PCa model, (B) Growth curve of PC3 PSMA+ tumors after indicated treatments. Data were analyzed using repeated‐measures ANOVA, (C) Representative BLI to show the tumor growth after indicated treatment, (D) Picture of isolated tumors and tumor weights after mice were sacrificed at endpoints. ( n = 5). Data were analyzed using the student t ‐test (two‐tailed), and (E) Immunofluorescent staining and quantitative analysis of Ki‐67 and TUNEL staining in the tumor tissues after the indicated treatment. Data were analyzed using the student t ‐test (two‐tailed). Scale bar, 50 µm. Data were shown as mean ± SD. **** P < 0.0001, *** P < 0.01, and ** P < 0.001; Abbreviation : NS, no significant.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: In Vivo, Isolation, Two Tailed Test, Staining, TUNEL Assay

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs@DM1 in combination with PTT exerts a combined anti‐tumor effect in the PSMA + PCa bone metastasis model. (A) Schematic illustration to show the design of animal experiments in the PCa bone metastasis model, (B) BLI to show the growth of bone metastatic tumor after indicated treatment, (C) Quantification of the fluorescence intensity in (B). Data were analyzed using the student t ‐test (two‐tailed), (D) Micro‐CT imaging of the tibia with a metastatic tumor. Tumor‐free BALB/c nude mice were used as normal, and (E) Bone mineral density of the tibia in different groups according to the result in (D). Data were analyzed using the student t ‐test (two‐tailed). Data were shown as mean ± SD. **** P < 0.0001, *** P < 0.05, ** P < 0.01, and * P < 0.001; Abbreviation : NS, no significant.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: Fluorescence, Two Tailed Test, Micro-CT, Imaging

Journal: Exploration

Article Title: PSMA‐Targeting Macrophage Membrane‐Coated Nanoparticles for Precision Diagnosis and Combination Therapy of Prostate Cancer

doi: 10.1002/EXP.20240393

Figure Lengend Snippet: P‐MMCNPs inhibit the growth of PSMA + tumors through macrophage‐associated immunomodulation in vivo. (A) Immunofluorescent staining and quantitative analysis of M1 and M2 macrophages in tumor tissues after P‐MMCNPs treatment. Data were analyzed using the student t ‐test (two‐tailed). Scale bar, 20 µm and (B) ELISA to examine the levels of different cytokines in tumor tissues. Data were analyzed using the student t ‐test (two‐tailed). Data were shown as mean ± SD. **** P < 0.0001, ** P < 0.05, and * P < 0.01.

Article Snippet: Then, the recombinant PSMA protein with His tag (Sino Biological, 15877‐H07H, China) was added to the plate for incubation.

Techniques: In Vivo, Staining, Two Tailed Test, Enzyme-linked Immunosorbent Assay