Platinum-perovskite nanocomposite-based Exosensor for specific detection of prostate cancer in clinical settings.

Exosomes, extracellular vesicles (EVs) with an average size of 50-150 nm, transfer various biomolecules and exchange signaling molecules between cells in a paracrine manner. Molecular investigations have revealed that EVs can reflect real-time metabolic changes in normal- and cancer-origin cells and thus harbor valid diagnostic biomarkers. Despite these advantages, the detection of low concentrations of cancer cell EVs in biological fluids is still a great challenge. Here, a new electrochemical Exosensor based on platinum-perovskite is developed for the direct detection of EVs using a biotinylated monoclonal CD63 antibody as a capture element. The label-free method exhibited higher sensitivity with a lower limit of quantification of 2000 EVs/µL with a dynamic linear range (LDR) of 2000 to 14,000 EVs/µL compared with other available methods. To enhance the selectivity of detection, EVs were simultaneously sandwiched between secondary antibodies of PSA (prostate-specific antigen), as an FDA-approved prostate cancer biomarker. Data indicated that this Exosensor can distinguish normal and cancer EVs in samples from healthy individuals and prostate cancer patients. Taken together, this technology offers a unique approach to label-free quantification of EVs and cancer detection in the early stages.

A smartphone-based immunochromatographic strip platform for on-site quantitative detection of antigenic targets.

To report the testing signal of an immunochromatographic assay for on-site quantitative detection, a portable and user-friendly smartphone-based biosensing platform is developed in this study. This innovative system is composed of an ambient light sensor inherent smartphone reader and a 3D-printed handhold device, a quantitative tool capable of directly interpreting carbon nanoparticle (CNP)-conjugated immunochromatographic strips. To showcase the platform capability, the smartphone-based immunochromatography system (SPICS) reader and device were successfully used in CNP strips for rapid detection of the early pregnancy marker human chorionic gonadotropin in female urine (HCG; limit of detection [LOD]: 0.30 mIU mL-1), prostate-specific antigen in patient blood (PSA; LOD: 0.28 ng mL-1) and ampicillin residue in animal milk (AMP; LOD: 0.23 ng mL-1). The results were fully correlated with conventional commercial instruments (R2 = 0.99). The SPICS platform exhibits significant advantages, including portability, cost-effectiveness, easy operation, and rapid and quantitative detection, making it a valuable on-site diagnosis tool for use in home and community healthcare facilities.

Long-term follow up of patients treated with a DNA vaccine (pTVG-hp) for PSA-recurrent prostate cancer.

We have previously reported two single-agent phase I trials, evaluating the dose or schedule, of a DNA vaccine (pTVG-HP) encoding prostatic acid phosphatase (PAP) administered with GM-CSF as the adjuvant. These were in patients with PSA-recurrent, radiographically non-metastatic, prostate cancer (PCa). We report here the long-term safety and overall survival of these patients. Specifically, 22 patients with non-metastatic, castration-sensitive PCa (nmCSPC) were treated with pTVG-HP, 100-1500 µg, administered over 12 weeks and followed for 15 y. 17 patients with non-metastatic castration-resistant PCa (nmCRPC) were treated with 100 µg pTVG-HP with different schedules of administration over 1 y and followed for 5 y. No adverse events were detected in long-term follow-up from either trial that were deemed possibly related to vaccination. Patients with nmCSPC had a median overall survival of 12.3 y, with 5/22 (23%) alive at 15 y. 8/22 (36%) died due to prostate cancer with a median survival of 11.0 y, and 9/22 (41%) died of other causes. Patients with nmCRPC had a median overall survival of 4.5 y, with 8/17 (47%) alive at 5 y. The presence of T-cells specific for the PAP target antigen was detectable in 6/10 (60%) individuals with nmCSPC, and 3/5 (60%) individuals with nmCRPC, many years after immunization. The detection of immune responses to the vaccine target years after immunization suggests durable immunity can be elicited in patients using a DNA vaccine encoding a tumor-associated antigen.Trial Registration: NCT00582140 and NCT00849121.

Immunological assay using a solid-state pore with a low limit of detection.

Emerging infectious diseases, cancer, and other diseases are quickly tested mainly via immune reactions based on specific molecular recognition between antigens and antibodies. By changing the diameter of solid-state pores, biomolecules of various sizes can be rapidly detected at the single-molecule level. The combination of immunoreactions and solid-state pores paves the way for an efficient testing method with high specificity and sensitivity. The challenge in developing this method is achieving quantitative analysis using solid-state pores. Here, we demonstrate a method with a low limit of detection for testing tumor markers using a combination of immunoreactions and solid-state pore technology. Quantitative analysis of the mixing ratio of two and three beads with different diameters was achieved with an error rate of up to 4.7%. The hybrid solid-state pore and immunoreaction methods with prostate-specific antigen (PSA) and anti-PSA antibody-modified beads achieved a detection limit of 24.9 fM PSA in 30 min. The hybrid solid-state pore and immunoreaction enabled the rapid development of easy-to-use tests with lower limit of detection and greater throughput than commercially available immunoassay for point-of-care testing.

Development of a first-in-class antibody and a specific assay for α-1,6-fucosylated prostate-specific antigen.

Prostate-specific antigen (PSA) levels are widely used to screen for prostate cancer, yet the test has poor sensitivity, specificity and predictive value, which leads to overdiagnosis and overtreatment. Alterations in the glycosylation status of PSA, including fucosylation, may offer scope for an improved biomarker. We sought to generate a monoclonal antibody (mAb) targeting α-1,6-fucosylated PSA (fuc-PSA) and to develop a tissue-based immunological assay for fuc-PSA detection. Immunogens representing fuc-PSA were used for immunisation and resultant mAbs were extensively characterised. The mAbs reacted specifically with fuc-PSA-specific glycopeptide, but not with aglycosylated PSA or glycan without the PSA peptide. Reactivity was confirmed using high-throughput surface plasmon resonance spectroscopy. X-ray crystallography investigations showed that the mAbs bound to an α-helical form of the peptide, whereas the native PSA epitope is linear. Protein unfolding was required for detection of fuc-PSA in patient samples. Peptide inhibition of fuc-PSA mAbs was observed with positive screening reagents, and target epitope specificity was observed in formalin-fixed, paraffin-embedded tissue samples. This research introduces a well-characterised, first-in-class antibody targeting fuc-PSA and presents the first crystal structure of an antibody demonstrating glycosylation-specific binding to a peptide.

An electrochemical/SERS dual-mode immunosensor using TMB/Au nanotag and Au@2D-MoS2 modified screen-printed electrode for sensitive detection of prostate cancer biomarker.

This study describes a novel dual-mode immunosensor that combines electrochemical (EC) and surface-enhanced Raman scattering (SERS) techniques for the detection of prostate-specific antigen (PSA), a biomarker associated with prostate cancer. The sensor consists of a nanocomposite of gold nanoparticles (AuNPs) deposited on two-dimensional (2D) molybdenum disulfide (Au@MoS2) modified on a working carbon electrode of a screen-printed electrode (SPE). Subsequently, the primary antibody (Ab1) is immobilized on the modified electrode, creating Ab1/Au@MoS2/SPE for specific recognition of the target PSA. In parallel, AuNPs are conjugated with a secondary antibody (Ab2) and a probe molecule, 3,3',5,5'-tetramethylbenzidine (TMB), leading nanotags (TMB/Ab2/AuNPs) formation exhibiting strong SERS and EC responses. Upon the presence of the target, sandwich immunocomplexes can be formed through antigen-antibody interactions (Ab1-PSA-Ab2). The differential pulse voltammetry (DPV) technique is employed for EC detection mode, while a handheld Raman spectrometer with a 785 nm excitation laser is utilized to collect SERS signals. The developed system demonstrates excellent selectivity and sensitivity, with low limits of detection (LODs) of 3.58 pg mL-1 and 4.83 pg mL-1 for EC and SERS sensing, respectively. Importantly, the dual-mode immunosensor proves effective quantifying PSA protein in human serum samples with good recovery. Given its high sensitivity and proficiency in analyzing biological samples, this proposed immunosensor holds promise as an alternative tool for the early diagnosis of cancers.

Development of a bioluminescent homogenous nanobody-based immunoassay for the detection of prostate-specific antigen (PSA).

Prostate cancer is the most prevalent cancer in men. At present, the diagnosis and screening of prostate cancer rely on the essential biomarker known as prostate-specific antigen (PSA). The main purpose of this study was to develop a novel immunoassay for the detection of PSA based on a tri-part split-nanoluciferase system and a nanobody targeting PSA. In our approach, two small components of the split-nanoluciferase, referred to as ß9 and ß10, were individually fused to two anti-PSA nanobodies, N7 and N23. When these proteins bind to PSA and in the presence of the third nanoluciferase component, called Δ11S, the split-nanoluciferase components are brought into close proximity, facilitating the reassembly of the active nanoluciferase and activation of luminescence. These proteins were expressed in a bacterial expression system, purified, and employed for the intended immunoassay. The developed immunoassay demonstrated the capability to sensitively detect PSA within a linear range from 1.0 to 20.0 ng/mL with LOD of 0.4 ng/mL, and the results obtained through this immunoassay agreed with those derived from the ELISA. Our study indicates that the homogeneous immunoassay developed with nanobodies exhibits remarkable specificity for PSA and can serve as a reliable, fast, and user-friendly test for detecting PSA.

Towards a Precision Medicine Approach and In Situ Vaccination against Prostate Cancer by PSMA-Retargeted oHSV.

Prostate specific membrane antigen (PSMA) is a specific high frequency cell surface marker of prostate cancers. Theranostic approaches targeting PSMA show no major adverse effects and rule out off-tumor toxicity. A PSMA-retargeted oHSV (R-405) was generated which both infected and was cytotoxic exclusively for PSMA-positive cells, including human prostate cancer LNCaP and 22Rv1 cells, and spared PSMA-negative cells. R-405 in vivo efficacy against LLC1-PSMA and Renca-PSMA tumors consisted of inhibiting primary tumor growth, establishing long-term T immune response, immune heating of the microenvironment, de-repression of the anti-tumor immune phenotype, and sensitization to checkpoint blockade. The in situ vaccination protected from distant challenge tumors, both PSMA-positive and PSMA-negative, implying that it was addressed also to LLC1 tumor antigens. PSMA-retargeted oHSVs are a precision medicine tool worth being additionally investigated in the immunotherapeutic and in situ vaccination landscape against prostate cancers.

Telomerase-based GX301 cancer vaccine in patients with metastatic castration-resistant prostate cancer: a randomized phase II trial.

Debate is around the optimal immunization regimen for cancer vaccines since too intense vaccination schedules may exhaust reactive lymphocytes. GX301 is a telomerase-based cancer vaccine whose safety and immunological effects were tested in a phase I trial applying an eight administrations schedule. Main objective of this study was to comparatively analyse safety and immunological response to three GX301 regimens in metastatic castration-resistant prostate cancer patients with response/disease stability after docetaxel chemotherapy. This was a multicentre, randomized, parallel-group, open-label trial registered with EudraCT (2014-000095-26) and ClinicalTrials.gov (NCT02293707, 2014). Ninety-eight patients were randomized to receive either eight (regimen 1), four (regimen 2) or two (regimen 3) vaccine administrations. Sixty-three patients were assessable for the primary immunological end-point. Vaccine-specific immune responses were evaluated by intracellular staining for IFN, elispot and cytotoxic assay at 90 and 180 days from baseline. No major side effects were recorded. A 54% overall immune responder rate was observed with 95% of patients showing at least one vaccine-specific immune response. Rate of immunological responders and number of immunizations were proportionally related, suggesting superiority of regimens 1 and 2 over regimen 3. Overall survival did not differ among regimens in both immunological responders and non-responders and was inversely associated (P = 0.002) with increase in the number of circulating CD8 + T regulatory cells at 180 days. These data indicate that GX301 cancer vaccine is safe and immunogenic in metastatic castration-resistant prostate cancer patients. Schedules with high number of administrations should be preferred in future studies due to their better immunological outcome.

Analysis of allergen profile in patients sensitized to canine allergen and potential Can f 5 cross-reactivity with human PSA.

Can f 5 allergy and possible cross-reactivity with human semen in which there are significant amounts of prostate-specific antigen (PSA) are particularly interesting aspects of allergy to dog. The objective of the study was to confirm cross-reactivity between human PSA and Can f 5 in a study of canine sensitised women. A total of 100 women (aged 18-73, 41 on average) with a positive history of animal fur allergy or positive skin prick tests to canine allergens were selected. Levels of Immunoglobulin E (IgE) specific to Can f 1, Can f 2, Can f 3, Can f 5 were determined. Patients with increased concentration of sIgE Can f 5 were selected for further inhibition testing using polystyrene microplate ELISA test coated with human PSA. In the studied population, allergy to Can f 5 dominated (52.3% of patients with increased concentration of canine-specific IgE were allergic to this allergenic component). In all analyzed cases, the concentration of IgE Can f 5 decreased after incubation on the ELISA plate coated with human PSA. The minimum decrease in concentration was 10.44%, the maximum was 37.73%, the average decrease was 21.6%. No statistically significant influence of the presence or absence of allergenic sIgE Can f 5 in blood serum on the occurrence of symptoms after intercourse was found. The study confirmed the moderate ability of Can f 5 to cross-react with human PSA sIgE, which may be clinically significant in some women. At the same time, symptoms of an allergy to male semen do not constitute a typical clinical presentation of allergy to Can f 5.

Phase I study of a multitargeted recombinant Ad5 PSA/MUC-1/brachyury-based immunotherapy vaccine in patients with metastatic castration-resistant prostate cancer (mCRPC).

BACKGROUND: Antitumor vaccines targeting tumor-associated antigens (TAAs) can generate antitumor immune response. A novel vaccine platform using adenovirus 5 (Ad5) vectors [E1-, E2b-] targeting three TAAs-prostate-specific antigen (PSA), brachyury, and MUC-1-has been developed. Both brachyury and the C-terminus of MUC-1 are overexpressed in metastatic castration-resistant prostate cancer (mCRPC) and have been shown to play an important role in resistance to chemotherapy, epithelial-mesenchymal transition, and metastasis. The transgenes for PSA, brachyury, and MUC-1 all contain epitope modifications for the expression of CD8+ T-cell enhancer agonist epitopes. We report here the first-in-human trial of this vaccine platform. METHODS: Patients with mCRPC were given concurrently three vaccines targeting PSA, brachyury, and MUC-1 at 5×1011 viral particles (VP) each, subcutaneously every 3 weeks for a maximum of three doses (dose de-escalation cohort), followed by a booster vaccine every 8 weeks for 1 year (dose-expansion cohort only). The primary objective was to determine the safety and the recommended phase II dose. Immune assays and clinical responses were evaluated. RESULTS: Eighteen patients with mCRPC were enrolled between July 2018 and September 2019 and received at least one vaccination. Median PSA was 25.58 ng/mL (range, 0.65-1006 ng/mL). The vaccine was tolerable and safe, and no grade >3 treatment-related adverse events or dose-limiting toxicities (DLTs) were observed. One patient had a partial response, while five patients had confirmed PSA decline and five had stable disease for >6 months. Median progression-free survival was 22 weeks (95% CI: 19.1 to 34). Seventeen (100%) of 17 patients mounted T-cell responses to at least one TAA, whereras 8 (47%) of 17 patients mounted immune responses to all three TAAs. Multifunctional T-cell responses to PSA, MUC-1, and brachyury were also detected after vaccination in the majority of the patients. CONCLUSIONS: Ad5 PSA/MUC-1/brachyury vaccine is well tolerated. The primary end points were met and there were no DLTs. The recommended phase II dose is 5×1011 VP. The vaccine demonstrated clinical activity, including one partial response and confirmed PSA responses in five patients. Three patients with prolonged PSA responses received palliative radiation therapy. Further research is needed to evaluate the clinical benefit and immunogenicity of this vaccine in combination with other immuno-oncology agents and/or palliative radiation therapy. TRIAL REGISTRATION NUMBER: NCT03481816.

PSA-Targeted Alpha-, Beta-, and Positron-Emitting Immunotheranostics in Murine Prostate Cancer Models and Nonhuman Primates.

PURPOSE: Most patients with prostate cancer treated with androgen receptor (AR) signaling inhibitors develop therapeutic resistance due to restoration of AR functionality. Thus, there is a critical need for novel treatment approaches. Here we investigate the theranostic potential of hu5A10, a humanized mAb specifically targeting free PSA (KLK3). EXPERIMENTAL DESIGN: LNCaP-AR (LNCaP with overexpression of wildtype AR) xenografts (NSG mice) and KLK3_Hi-Myc transgenic mice were imaged with 89Zr- or treated with 90Y- or 225Ac-labeled hu5A10; biodistribution and subcellular localization were analyzed by gamma counting, PET, autoradiography, and microscopy. Therapeutic efficacy of [225Ac]hu5A10 and [90Y]hu5A10 in LNCaP-AR tumors was assessed by tumor volume measurements, time to nadir (TTN), time to progression (TTP), and survival. Pharmacokinetics of [89Zr]hu5A10 in nonhuman primates (NHP) were determined using PET. RESULTS: Biodistribution of radiolabeled hu5A10 constructs was comparable in different mouse models. Specific tumor uptake increased over time and correlated with PSA expression. Treatment with [90Y]/[225Ac]hu5A10 effectively reduced tumor burden and prolonged survival (P ≤ 0.0054). Effects of [90Y]hu5A10 were more immediate than [225Ac]hu5A10 (TTN, P < 0.0001) but less sustained (TTP, P < 0.0001). Complete responses were observed in 7 of 18 [225Ac]hu5A10 and 1 of 9 mice [90Y]hu5A10. Pharmacokinetics of [89Zr]hu5A10 were consistent between NHPs and comparable with those in mice. [89Zr]hu5A10-PET visualized the NHP-prostate over the 2-week observation period. CONCLUSIONS: We present a complete preclinical evaluation of radiolabeled hu5A10 in mouse prostate cancer models and NHPs, and establish hu5A10 as a new theranostic agent that allows highly specific and effective downstream targeting of AR in PSA-expressing tissue. Our data support the clinical translation of radiolabeled hu5A10 for treating prostate cancer.

Utility of a Fifth-Generation Ultrasensitive Prostate-Specific Antigen Assay for Monitoring Prostate Cancer Patients after Radical Prostatectomy with 3 Years of Follow-Up.

BACKGROUND: We investigated an ultrasensitive prostate-specific antigen (uPSA) immunoassay (MesoScale; lower limit of detection (LLD) of 0.0035 pg/mL) to monitor patients with prostate cancer (PCa) following radical prostatectomy (RP) and to examine whether changes in PSA in the conventionally undetectable range (<1 pg/mL) can predict biochemical relapse (BCR). METHODS: We measured uPSA in serial serum samples (N = 100) collected from 20 RP cases with a third-generation ELISA (LLD of 1 pg/mL) and the fifth-generation MesoScale assay. We analyzed the PSA nadir changes to classify patients into BCR or non-BCR groups, observed the trends in PSA kinetics, and associated BCR status with clinicohistopathological features. RESULTS: The ELISA could quantify PSA in only 38% of the RP samples, detecting BCR in 7 of 20 patients with PCa. The MesoScale assay quantified PSA in all samples, showing 8 of 20 patients with BCR. However, there was no significant difference between the median time to BCR detection based on ELISA (1016 days) compared with MesoScale data (949 days). Gleason scores were higher in the BCR groups compared with non-BCR. There was no significant difference for other clinicohistopathological parameters. CONCLUSIONS: The uPSA MesoScale technology could track miniscule changes in serum PSA in the range of 0.003-1 pg/mL in all RP cases. However, PSA kinetics and nadir at concentrations <2 pg/mL fluctuated, and increases below this range could not reliably suggest signs of BCR. Instead, ultrasensitive fifth-generation PSA assays may hold clinical potential for measuring the low concentrations of PSA in women for various medical contexts.

Systemic Inflammatory Markers Are Predictive of the Response to Brachytherapy in the Prostate.

We analyzed the influence of the neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) on the biochemical recurrence (BCR) in low-intermediate risk prostate cancer (PCa). A total of 604 patients treated with exclusive brachytherapy for low- and intermediate-risk cancers were included in this study. No patient received either androgen deprivation or brachytherapy as a boost. BCR was defined according to the Phoenix definition (nadir prostatic specific antigen (PSA) +2). The median follow-up was 60 months (IQR 44-48 months). An NLR > 3 was more frequent in statin users (p = 0.025), but not in diabetics (p = 0.079). In univariate analysis (UVA) and multivariate analysis (MVA), a NLR > 3 (MVA p = 0.03), as well as Cancer of the Prostate Risk Assessment (CAPRA) low- vs. intermediate-risk (MVA p = 0.04), were predictive of BCR. When combining the NLR score with the CAPRA risk group, CAPRA intermediate risk patients with an NLR ≤ 3 (n = 157) had the worst (p = 0.0276) BCR rates, with a 5-year recurrence-free survival (p = 0.004, Bonferroni correction for six comparisons p = 0.024). We were able to identify a subgroup of PCa patients with CAPRA intermediate-risk and an NLR ≤ 3 who had worse BCR. This is in contrast to most other cancers, which have a worse prognosis when the NLR is high.

Upconversion luminescence-infrared absorption nanoprobes for the detection of prostate-specific antigen.

Aiming to the ongoing challenge of accurate and sensitive detection for cancer biomarkers, antibody-functionalized NaYF4:Yb3+, Er3+@SiO2 nanorods were developed as upconversion luminescence (UCL)-infrared absorption (IRA) nanoprobes. Benefiting from the shielding effect of the SiO2 shell, an enhanced UCL was achieved. Additionally, an IRA detection signal was introduced by the Si-O-Si bonds of SiO2. Its mutual verification with UCL signal was favorable for ensuring the accuracy of the assay. A UCL-IRA sandwich detection method was established for the detection of the prostate-specific antigen. The UCL intensity at 542 nm and IRA at 1095 cm-1 were chosen for quantitative assay. The method has high sensitivity (0.05 pg mL-1) and selectivity. The range of detection (200 fg mL-1-200 ng mL-1) was singnificantly broadened compared with that of single-readout UCL or IRA detection. The assay performance of human serum samples demonstrated the practicability of the method in clinical cancer diagnosis. Graphical abstract.

Self-assembled biotin-phenylalanine nanoparticles for the signal amplification of surface plasmon resonance biosensors.

A strategy for amplifying the signal of surface plasmon resonance (SPR) biosensors is reported. Biotinylated phenylalanine (Biotin-Phe) monomers were rapidly self-assembled into nanoparticles in a mild environment. The self-assembled nanoparticles were then used as the carriers of streptavidin-antibody complexes by the streptavidin-biotin interaction. The signal was amplified because of the high molecular weight of the nanoparticle-streptavidin-antibody conjugate. With prostate-specific antigen as a model analyte, the target concentration as low as 1 pg mL-1 was readily measured. The results of the nanoparticle-enhanced SPR biosensor for analysis of serum samples are well consistent with those achieved by the enzyme-linked immunosorbent assays. This work is valuable for designing of various optical and electronic biosensors through the streptavidin-biotin interaction. Graphical abstract.

Electrochemical assays for determination of H2O2 and prostate-specific antigen based on a nanocomposite consisting of CeO2 nanoparticle-decorated MnO2 nanospheres.

A nanocomposite consisting of CeO2 nanoparticle-decorated MnO2 nanospheres (CeO2@MnO2) was synthesized for the first time via a hydrothermal method. CeO2@MnO2 was exploited to construct an electrochemical assays for detecting H2O2 and prostate-specific antigen (PSA) with square wave voltammetry (SWV). The electrochemical results proved that CeO2@MnO2 owned a better electrocatalytic effect towards H2O2 reduction than pure MnO2 NS and CeO2 NP due to the synergistic effect between MnO2 NS and CeO2 NP. Under optimized conditions, CeO2@MnO2-based assay can be applied to detect H2O2 in the range 1 to 3.0 × 103 µmol L-1. The label-free electrochemical immunoassay based on CeO2@MnO2 displayed linearly with concentrations of PSA from 0.005 to 50.0 ng mL-1. The electrochemical assays also possessed acceptable sensitivity, selectivity, and stability. The study showed that CeO2@MnO2 hold great potential as a biosensing platform and the clinical determination of tumor markers in human serum. Graphical abstract A nanocomposite consisting of CeO2 nanoparticles decorated MnO2 nanospheres (CeO2 @MnO2) was firstly synthesized via a hydrothermal method. CeO2@MnO2 was firstly exploited to construct electrochemical assays for detecting H2O2 and prostate-specific antigen (PSA) with square wave voltammetry (SWV), respectively. The electrochemical results proved that CeO2@MnO2 owned better electrocatalysis towards H2O2 reduction than pure MnO2 NS and CeO2 NP due to the synergistic effect between MnO2 NS and CeO2 NP. Under optimized conditions, CeO2@MnO2 based assay relative to the H2O2 system can be applied to detect H2O2 with range from 1 to 3.0 × 103 µmol L-1. The label-free electrochemical immunoassay based on CeO2@MnO2 relative to the H2O2 system displayed linearly with concentrations of PSA from 0.005 to 50.0 ng mL-1. The electrochemical assays also possessed acceptable sensitivity, selectivity and stability. The study showed that CeO2@MnO2 hold great potential for biosensing platform and the clinic determination of tumor markers in human serum.

Development of an optical microfiber immunosensor for prostate specific antigen analysis using a high-order-diffraction long period grating.

Fiber-optic biosensors are of great interest to many bio/chemical sensing applications. In this study, we demonstrate a high-order-diffraction long period grating (HOD-LPG) for the detection of prostate specific antigen (PSA). A HOD-LPG with a period number of less than ten and an elongated grating pitch could realize a temperature-insensitive and bending-independent biosensor. The bio-functionalized HOD-LPG was capable of detecting PSA in phosphate buffered saline with concentrations ranging from 5 to 500 ng/ml and exhibited excellent specificity. A limit of detection of 9.9 ng/ml was achieved, which is promising for analysis of the prostate specific antigen.

In Pursuit of Stability Enhancement of a Prostate Cancer Targeting Antibody Derived from a Transgenic Animal Platform.

Accelerated timelines necessitate the discovery of fully human antibodies as biotherapeutics using transgenic animals with a notion that such mAbs bypass humanization. A transgenic animal derived mAb (PCa75) targeted against a prostate cancer antigen had several 'unusual residues' (rare somatic hypermutations, rSHM, with positional frequency of <1%) that resulted in compromised biophysical properties (Tm = 61 °C and intrinsic stability ΔGu = 24.3 kJ/mol) and a sub-optimal immunogenicity profile. In our quest for quality medicine, we pursued antibody engineering strategies to enhance the stability of PCa75. PCa62, an engineered variant of PCa75, retained function while significantly improving the drug-like attributes of the molecule (Tm = 75 °C and intrinsic stability ΔGu = 63.5 kJ/mol). rSHM is rather prevalent, 18 out the 21 approved transgenic animal-derived antibodies have at least one 'unusual residue'. Thus, engineering of rSHM remains critical to enhance the stability and minimize immunogenicity risk of biotherapeutics.

Genetic signature of prostate cancer mouse models resistant to optimized hK2 targeted α-particle therapy.

Hu11B6 is a monoclonal antibody that internalizes in cells expressing androgen receptor (AR)-regulated prostate-specific enzyme human kallikrein-related peptidase 2 (hK2; KLK2). In multiple rodent models, Actinium-225-labeled hu11B6-IgG1 ([225Ac]hu11B6-IgG1) has shown promising treatment efficacy. In the present study, we investigated options to enhance and optimize [225Ac]hu11B6 treatment. First, we evaluated the possibility of exploiting IgG3, the IgG subclass with superior activation of complement and ability to mediate FC-γ-receptor binding, for immunotherapeutically enhanced hK2 targeted α-radioimmunotherapy. Second, we compared the therapeutic efficacy of a single high activity vs. fractionated activity. Finally, we used RNA sequencing to analyze the genomic signatures of prostate cancer that progressed after targeted α-therapy. [225Ac]hu11B6-IgG3 was a functionally enhanced alternative to [225Ac]hu11B6-IgG1 but offered no improvement of therapeutic efficacy. Progression-free survival was slightly increased with a single high activity compared to fractionated activity. Tumor-free animals succumbing after treatment revealed no evidence of treatment-associated toxicity. In addition to up-regulation of canonical aggressive prostate cancer genes, such as MMP7, ETV1, NTS, and SCHLAP1, we also noted a significant decrease in both KLK3 (prostate-specific antigen ) and FOLH1 (prostate-specific membrane antigen) but not in AR and KLK2, demonstrating efficacy of sequential [225Ac]hu11B6 in a mouse model.