Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: List of aptamers used in this study.

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: In Vitro, Inhibition, Activity Assay

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Fluorescence microscopy. ( A ) Direct fluorescence method. FAM labeled anti-PSMA RNA aptamer (A9g) was incubated with either PC3(PSMA+) or PC3(PSMA-) cells. A high salt wash step was performed to remove unbound or surface bound RNA. Internalized RNA was visualized using fluorescence microscopy. A scrambled, non-internalizing aptamer (Scr) was used as a negative control in these experiments. Florescence images were overlaid with DAPI and P/C (phase contrast) channels. Arrows indicate perinuclear localization of internalized A9g aptamer. ( B ) Antibody amplification method. FAM-labeled anti-TrkB RNA aptamer (C4-3) was incubated with either TrkB expressing or non-expressing HEK293 cells at 37 °C. FAM-labeled control aptamer (Scr) was used as a control for specificity. Unbound and surface bound RNA was removed as described above. Internalized RNA signal was amplified by incubation with an anti-FITC antibody and Alexa488 secondary antibody. Vehicle treated cells (No RNA) or cells subjected to incubation with antibodies alone (Ab control) were used as controls.

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Fluorescence, Microscopy, Labeling, Incubation, Negative Control, Amplification, Expressing, Control

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Plate-reader assay to assess aptamer binding and internalization. For the binding experiments, cells were fixed to inhibit active transport before incubation with the RNA aptamers. Live cells were used for the internalization experiments. ( A ) Binding ( left ) and internalization ( right ) of A9g into PSMA-expressing prostate cancer cells. ( B ) Binding ( left ) and internalization ( right ) of E1 aptamer into rat HER2-expressing mammary carcinoma cells. Cells only (no RNA) controls were carried out for each condition. Fluorescence was measured using an Analyst HT plate reader. (*, p < 0.001).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Incubation, Expressing, Fluorescence

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Evaluation of aptamer binding and internalization by flow cytometry. ( A ) Cell-specific binding of a human HER2 aptamer-Qdot conjugate. Cell lines expressing HER2 receptor (N202.1E-hHER2 and SKBR3) and HER2 non-expressing cell line (N202.1E) were incubated for 45min at 37 °C with human HER2 aptamer conjugated to Qdots (605nm). Cell-specific aptamer binding was evaluated by flow cytometry (upper panel). Quantification of specific fluorescence signal is shown in the middle panel bar graph. Cell surface human HER2 receptor expression in N202.1E, N202.1E(hHER2) and SKBR-3 cells (grey for isotype control, colored histograms for anti-HER2 Ab) (lower panel). ( B ) Measurement of A9g cell-internalization. PSMA-positive cells were incubated with FAM-A9g aptamer for 30 min at either 4 °C (left top panel) or 37 °C (right top panel). Cells were then washed with either DPBS or a High Salt (DPBS plus 0.5M NaCl) wash for 5min at 4 °C. The high salt wash step removes any unbound or surface bound aptamer. Bound and/or internalized aptamers were subsequently visualized using flow cytometry. *, internalized aptamer fraction (middle right panel). Fluorescence intensity quantified in the bar graph (**, p < 0.005).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Flow Cytometry, Expressing, Incubation, Fluorescence, Control

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Quantitative and ultrasensitive internalization method ( “QUSIM” ). ( A ) 96-well microplate NIR Odyssey imager scan of serial dilutions for binding aptamer-NIR conjugate (A9g-NIR) and mutant, non-binding sequence conjugate (A9g.6-NIR) ( upper panel ) and standard curves with linear regression for RNA aptamer quantification ( lower panel ). ( B ) Quantification of the amount of aptamer-NIR internalized into PC3(PSMA+) cells vs. PC3(PSMA–) cells. ( C ) Time-dependent cell uptake of binding aptamer (A9g-NIR) vs. non-binding (mutant) aptamer (A9g.6-NIR). ( D ) Kinetics of specific A9g internalization using one-phase association curve fit (R 2 = 0.9924, k = 0.542 min −1 , half-time = 1.278 min).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Mutagenesis, Sequencing

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: RNA-RIP assay. (A) Schematic of A9g-saporin conjugate internalization and RIP effect leading to cell death. (B) Dose dependent response of A9g-saporin and control, A9g.6-saporin, conjugates in PC3(PSMA+) ( left ) and PC3(PSMA–) cells ( right ).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Control

Journal: bioRxiv

Article Title: Pooled screening identifies combinatorial CAR signaling domains for next-generation CAR-M immunotherapies

doi: 10.1101/2025.02.16.638489

Figure Lengend Snippet: (A) Schematic representation of the pro-phagocytic module screening strategy. (B) Correlation between log2 fold change and −log10 rho scores for pro-phagocytic CAR variant screens. Data represent 18 biological replicates across six NaSDC lentivirus batches. (C) FACS-based quantification of PC3-PSMA tumor cell phagocytosis by CAR-Ms (n = 4 biological replicates). (D) Schematic representation of the pro-inflammatory module screening strategy. (E) Correlation between log2 fold change and −log10 rho scores for pro-inflammatory CAR variant screens. Data represent 15 biological replicates across five NaSDC lentivirus batches. (F) Relative mRNA expression of TNF , IL6 and IL1B in CAR-Ms cocultured with PSMA positive target tumor cells (n = 4 biological replicates). (G) Schematic representation of the pro-infiltrating module screening strategy. (H) Correlation between log2 fold change and −log10 rho scores for pro-infiltrating CAR variant screens. Data represent 9 biological replicates across three NaSDC lentivirus batches. (I) Relative mRNA expression of MMP family genes in CAR-Ms cocultured with PSMA positive target tumor cells (n = 4 biological replicates). Dashed lines in ( C , F , I ) indicate −log10(0.05) (horizontal) and log2 fold change of truncated CARs (CARΔ) (vertical). Data in (C) are presented as mean ± s.e.m. and analyzed using one-way ANOVA followed by Dunnett’s multiple-comparison test.

Article Snippet: Biotinylated human PSMA proteins (ACROBiosystems) were added to the beads at a concentration sufficient to occupy 25% of the binding sites.

Techniques: Variant Assay, Expressing, Comparison

Journal: bioRxiv

Article Title: Pooled screening identifies combinatorial CAR signaling domains for next-generation CAR-M immunotherapies

doi: 10.1101/2025.02.16.638489

Figure Lengend Snippet: (A) Description of CAR variants used for individual characterization. (B) FACS-based quantification of PC3-PSMA tumor cell phagocytosis by UTD and CAR-Ms (n = 4 biological replicates). (C) Flow cytometric histograms showing surface expression of CD80 and CD86 in UTD and CAR-Ms cocultured with PSMA positive tumor cells. (D) Flow cytometric histograms of reactive oxygen species (ROS) levels in UTD and CAR-Ms cocultured with PSMA positive tumor cells. DCF, 2’-7’dichlorofluorescein. (E) Relative mRNA expression of TNF and IL6 in UTD and CAR-Ms cocultured with PSMA positive tumor cells (n = 4 biological replicates). (F) Relative mRNA expression of MMP11 and MMP13 in UTD and CAR-Ms cocultured with PSMA positive tumor cells (n = 4 biological replicates). Data in ( B) are presented as mean ± s.e.m. and analyzed using one-way ANOVA followed by Dunnett’s multiple-comparison. Data in ( C , D) are representative of three independent experiments. Dashed lines ( C , D ) represent the mean fluorescence intensity (MFI) for UTD macrophages.

Article Snippet: Biotinylated human PSMA proteins (ACROBiosystems) were added to the beads at a concentration sufficient to occupy 25% of the binding sites.

Techniques: Expressing, Comparison, Fluorescence

Journal: bioRxiv

Article Title: Pooled screening identifies combinatorial CAR signaling domains for next-generation CAR-M immunotherapies

doi: 10.1101/2025.02.16.638489

Figure Lengend Snippet: (A) Schematic illustration of the humanized immune system TME model. PC3-PSMA tumors were established bilaterally, followed by PBMC engraftment on day 7. On day 14, 5 × 10 6 UTD or CAR-Ms were injected IT. Tumors were harvested on day 18 for scRNA-seq. SC, subcutaneously. IV, intravenously. IT, intratumorally. (B) Proportions of cell types from scRNA-seq (left) and tSNE-based clustering of all sequenced cells (right). B, B cell. T, T cell. Mac, macrophage. (C) Dimensionality reduction (tSNE) and clustering for macrophages. Data are colored by treatment conditions. (D) Density plots within tSNE maps depicting the distribution of CAR-expressing cells. (E) AUC scores of M1 and M2 gene sets in the indicated macrophage clusters. (F) Feature plots showing expression levels of selected genes. (G) MFI of HLA-ABC and HLA-DR in UTD and CAR-Ms after 24 h coculture with PSMA positive tumor cells (n = 3 biological replicates). (H) AUC scores of T cell cytotoxicity and exhaustion gene sets in the indicated T cell clusters. Unless specified otherwise, data are presented as mean ± s.e.m. and analyzed using the two-tailed Student’s t-test.

Article Snippet: Biotinylated human PSMA proteins (ACROBiosystems) were added to the beads at a concentration sufficient to occupy 25% of the binding sites.

Techniques: Injection, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Pooled screening identifies combinatorial CAR signaling domains for next-generation CAR-M immunotherapies

doi: 10.1101/2025.02.16.638489

Figure Lengend Snippet: (A) Specific killing of PC3-PSMA cells by UTD and COMB7 CAR-Ms at varying E/T ratios. Data represent two independent experiments from two human donors. (B) Specific killing of PC3-PSMA cells by UTD, CD247 CAR-Ms or COMB7 CAR-Ms at an E/T ratio of 3/1. Data collected from two human donors. (C-D) Specific killing of HER2 + SKOV3 or GPC3 + HepG2 cells by UTD or COMB7 CAR-Ms at an E/T ratio of 3/1 (n = 6 biological replicates). (E) NCG mice were IP injected with 1 × 10 6 PC3-PSMA cells, followed by treatment with PBS, 5 × 10 6 UTD, or 5 × 10 6 COMB7 CAR-Ms one day later. (F) Overall survival for mice in each treatment group (n = 6 for PBS, n = 5 for UTD and COMB7). (G) Tumor burden measured by bioluminescence (total flux). Data shown are individual values (n = 6 for PBS, n = 5 for UTD and COMB7). Unless specified otherwise, data are presented as mean ± s.e.m. and analyzed using the two-tailed Student’s t-test.

Article Snippet: Biotinylated human PSMA proteins (ACROBiosystems) were added to the beads at a concentration sufficient to occupy 25% of the binding sites.

Techniques: Injection, Two Tailed Test

Journal: Genome Medicine

Article Title: Evaluating the transcriptional fidelity of cancer models

doi: 10.1186/s13073-021-00888-w

Figure Lengend Snippet: Immunofluorescence of selected cell lines. a Classification profiles (left) and IF expression (middle) of Caov-4 (OV-positive control), HEC-59 (UCEC-positive control), and SK-OV-3 for WT1 (OV biomarker) and HOXB6 (uterine biomarker). The rightmost bar plots quantify the average percentage of positive cells for WT1 (top-right) and HOXB6 (bottom-right). b Classification profiles (left) and IF expression (middle) of Caov-4, NCCIT (germ cell tumor-positive control), and A2780 for WT1 and LIN28A (germ cell tumor biomarker). Classification was performed on replicates of NCCIT RNA-seq profiles. The rightmost bar plots quantify the average percentage of positive cells for WT1 (top-right) and LIN28A (bottom-right). c Classification profiles (left) and IF expression (middle) of Vcap (PRAD positive control), RT4 (BLCA-positive control), and PC-3 for FOLH1 (prostate biomarker) and PPARG (urothelial biomarker). The rightmost bar plots quantify the average percentage of positive cells for FOLH1 (top-right) and PPARG (bottom-right)

Article Snippet: Cells were incubated with primary antibodies to goat HOXB6 (10 μg/mL, PA5-37867, Invitrogen), mouse WT1 (10 μg/mL, MA1-46028, Invitrogen), rabbit PPARG (1:50, ABN1445, Millipore), mouse FOLH1 (10 μg/mL, UM570025, Origene), and rabbit LIN28A (1:50, #3978, Cell Signaling) in Antibody Diluent (S080981-2, DAKO), at 4 °C overnight followed with three 5 min washes in TBST.

Techniques: Immunofluorescence, Expressing, Positive Control, Biomarker Discovery, RNA Sequencing

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Establishment of monovalent aAVCs co-expressing CD1d and prostate tumor antigens for cancer therapy. (A) aAVCs expressing PSMA were created by electroporating NIH3T3 cells with PSMA and murine CD1d mRNA, followed by loading with α-galactosylceramide (α-GalCer). PSMA protein expression level was assessed using western blotting. (B) CD1d surface expression on aAVC-PSMA was assessed by flow cytometry (open, aAVC-PSMA; shaded, isotype). (C) aAVC-PSA was prepared as described in (A) , with PSA being transfected instead. PSA protein expression in aAVC-PSA was measured using western blot analysis. (D) CD1d surface expression on aAVC-PSA was assessed using flow cytometry (open, aAVC-PSA; shaded, isotype). (E) aAVC-PAP was prepared as described in (A) , with PAP transfected instead. The PAP protein in aAVC-PAP was measured using western blot analysis. (F) as similar to (A) , but CD1d surface expression on aAVC-PAP was assessed by flow cytometry (open, aAVC-PAP; shaded, isotype).

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Expressing, Western Blot, Flow Cytometry, Transfection

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: iNKT cell activation by aAVCs. (A, C, E) α-GalCer presentation on aAVC-PSMA (A) , aAVC-PSA (B) , and aAVC-PAP (C) . aAVCs were co-cultured with the Vα14 iNKT cell hybridoma 1.B2 for 24 h (upper), and IL-2 production in the culture supernatant was evaluated using IL-2 ELISA (n=4, mean ± SEM) (lower); ***P<0.001 (Mann-Whiteny). (B, D, F) C57BL/6 mice were injected intravenously with 5 × 10 5 aAVC-PSMA (B) , aAVC-PSA (D) , and aAVC-PAP (F) . Spleens were removed after 3 days, and splenic iNKT was analyzed after staining with CD19-FITC, CD1d-dimer + APC, and 7-AAD (upper). Representative dot plots showing the frequency of splenic iNKT cells (lower left) and summary(n=3, mean ± SEM) (lower right) were shown. **P<0.01, ***P<0.001(Mann-Whiteny).

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Injection, Staining

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Antitumor response by aAVCs in a prophylactic model. (A) Establishment of B16-PSMA (A) , B16-PSA (C) , and B16-PAP (E) cells. After introducing the PSMA, PSA, or PAP gene into B16, stable PSMA, PSA, or PAP protein expression was verified using western blot analysis. (B, D, F) Mice were immunized with or without 5 × 10 5 aAVC-PSMA, aAVC-PSA, or aAVC-PAP on day 0. Following this, mice were challenged with 5 × 10 5 B16-PSMA (B) , B16-PSA (D) , or B16-PAP (F) , respectively on day 14 (n=5 per group, mean ± SEM); *P<0.05 . *** P <0.001.

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Antitumor response by aAVCs in a therapeutic model. The antitumor response was evaluated in a therapeutic model in which the therapy was administered at a tumor volume of around 50 mm 3 . (A) Mice were inoculated with 5 × 10 5 B16-PSMA cells and treated with or without 5 × 10 5 aAVC-PSMA on day 12 (n=8 per group, mean ± SEM); * P <0.05. (B) Mice were inoculated with 5 × 10 5 B16-PSA cells and treated with or without 5 × 10 5 aAVC-PSA on day 7 (n=6 per group, mean ± SEM); *** P <0.001. (C) Mice were inoculated with 5 × 10 5 B16-PAP cells and treated with or without 5 × 10 5 aAVC-PAP on day 10 (n=7–8 per group, mean ± SEM); *** P <0.001.

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques:

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Induction of tumor epitope spreading by aAVC vaccination. (A, B) B16-PSMA/PSA/PAP cells were established using a retroviral vector expressing PSMA, PSA, or PAP, respectively. Tumor expression was verified using RT-PCR and western blotting. (C, E, G) Antitumor effect of various prostate antigen-expressing aAVC therapies. Mice were inoculated with 5 × 10 5 B16-PSMA/PSA/PAP cells and treated with or without 5 × 10 5 aAVC-PSMA (C) , aAVC-PSA (E) , or aAVC-PAP (G) on day 7 (n=5 per group, mean ± SEM); *P< 0.05. (D, F, H) Epitope spread of prostate antigens in CD8 + T cells after aAVC therapy. Ten days after vaccination with aAVC-PSMA (B) , aAVC-PSA (D) , or aAVC-PAP (F) , PSMA-, PSA-, or PAP-specific CD8 + T cells were analyzed using an IFN-γ ELISPOT assay. For this, splenic CD8 + T cells isolated from the immunized mice were cultured with splenic CD11c + dendritic cells (DCs) from naïve mice that had been cultured in the presence or absence of PSMA, PSA, or PAP-PepTivator for 24 h. (n=4 per group, mean ± SEM); *P< 0.05; ***P< 0.001; ns : not significant, according to Tukey’s test.

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Retroviral, Plasmid Preparation, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunospot, Isolation, Cell Culture

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Establishment of a multivalent aAVC and prostate cancer antigen-specific CD8 + T cell response. (A) Establishment of multivalent artificial adjuvant vector cells for the prostate (aAVC-PROS). Three prostate cancer antigen-expressing artificial adjuvant cells (aAVC-PSMA/PSA/PAP and aAVC-PROS) were established by co-transfecting NIH3T3 cells with PSMA, PSA, PAP, and murine CD1d mRNA and then loaded with α-GalCer. (B) Tumor protein antigen in aAVC-PROS. The amount of PSMA, PSA, and PAP was determined using western blot analysis. (C) The prostate antigen-specific CD8 + T cell response induced by aAVC. Mice were intravenously immunized with 5 × 10 5 aAVC-PROS. After one week, PSMA-, PSA-, or PAP-specific CD8 + T cells were analyzed using an IFN-γ ELISPOT assay. Splenic CD8 + T cells isolated from the immunized mice were cultured with splenic CD11c + DCs from naïve mice that had been cultured in the presence or absence of PSMA-, PSA-, or PAP-PepTivator for 24 h. (n=4 per group, mean ± SEM) *P<0.05 ; ***P<0.001 ; ns : not significant, according to Tukey’s test.

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Adjuvant, Plasmid Preparation, Expressing, Western Blot, Enzyme-linked Immunospot, Isolation, Cell Culture

Journal: Frontiers in Immunology

Article Title: Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ

doi: 10.3389/fimmu.2024.1345037

Figure Lengend Snippet: Comparison of the antitumor effect of divalent and trivalent prostate antigen-expressing aAVCs. Divalent artificial adjuvant cells (aAVC-PSMA/PSA) were established by co-transfecting NIH3T3 cells with PSMA, PSA, and murine CD1d mRNA. (A) Mice were inoculated with 5 × 10 5 B16-PSMA/PSA/PAP cells and treated with or without 5 × 10 5 divalent aAVC-PSMA/PSA or trivalent aAVC-PROS on day 7. (B) Comparison of the antitumor effects of monovalent (aAVC-PSMA, aAVC-PSA, or aAVC-PAP), divalent (aAVC-PSMA/PSA), or trivalent aAVCs (aAVC-PROS). The tumor size was evaluated by comparing untreated mice with treated mice on days 10 (Early phase) and 19 (Late phase). *Plt;0.05, **P<0.01, ***P<0.001; ns: Tukey’s test.

Article Snippet: PepTivator PSMA (#130-099-795), PepTivator PSA (#130-099-800), and PepTivator PAP (#130-096-767) were purchased from Miltenyi Biotech Inc.

Techniques: Comparison, Expressing, Adjuvant