Analyte Concentration Research Articles

Fabrication of mechanically alloyed high entropy alloys (20-Fe-20Cr-20Ni-20Mn-20Ti) modified carbon paste electrode sensor for the cyclic voltammetric determination of methyl orange

We have successfully fabricated high entropy alloys (HEA) of composition 20-Fe-20Cr-20Ni-20Mn-20Ti by ball milling method for 20 h after optimizing the milling parameters. Generally, the HEAs are very popular for their multi-elemental cocktail effect and exhibit excellent high temperature oxidation resistance, high strength, electromagnetic, wear and corrosion resistance properties. But, HEAs also exhibit significant electrocatalytic properties due to their unique morphology, composition, and structure. Therefore, we have used planetary ball milled HEA powders for the electrochemical determination of methyl orange (MO). MO is recalcitrant and very difficult to degrade and can affect the humans as well as the environment. Therefore, we must determine the presence of MO. We have investigated the effect of scan rate, modifier and analyte concentration, and the pH variation on the anodic peak current (Ipa) of MO. The calculated active syrface area of bare carbon paste electrode (BCPE) and HEA-modified carbon paste electrode (HEA-MCPE) were found to be 0.041 cm2 and 0.137 cm2 respectively. We have obtained the maximum oxidation peak current for the 8 mg HEA-MCPE at a pH of 7.2 during the electrochemical oxidation of MO. The increased Ipa in basic medium is due to structural transformation of MO from quinoid form (in acidic medium) to azo structure (in basic medium). We have also calculated the number of electrons and the protons involved in the electrochemical reaction. The calculated values of detection limit (DL) and quantification limit (QL) of 8 mg HEA-MCPE was found to be 0.114 × 10−8 M and 0.38 × 10−8 M respectively.

Chromolaena Odorata Leaves Derived Carbon Dot‐Based Fluorescence/Second‐Order Scattering Ratiometric Sensor for Fe3+ and Vitamin C

Herein, a ratiometric fluorescence and second‐order scattering sensor with plant leaf‐derived carbon dots (CDs) for Fe3+ and vitamin C is reported. The CDs are prepared from Chromolaena odorata leaves through a one‐pot hydrothermal method. The synthesized CDs have a quantum yield of 22.6%. The spherical dots have an average size of 3 nm and the chemical composition is found as nitrogen (7%) being the highest heteroatom present along with carbon and oxygen atoms in it. The ratio between fluorescence intensity at 445 nm and second‐order scattering constitutes the sensor output. A linear relationship is found between the output and analyte concentration in the range 0–70 and 0–10 μM with limit of detection 0.12 and 1.2 μM for Fe3+ and vitamin C, respectively. Static mechanism along with inner filter effect has been identified as the underlying quenching mechanism for Fe3+ activity. The strong chelation of Fe3+ by vitamin C further enhances the fluorescence. The change in the second‐order scattering intensity is explained by the formation and disruption of CDs/Fe3+ complex. The sensor also shows satisfactory performance with borewell water (Fe3+) and commercial tablet (vitamin C) with recovery rate ranging between 98.6% and 103.5%.

Multilingual pre-service teacher's knowledge and understanding of analytical geometry content

Multilingual pre-service teacher's knowledge and understanding of analytical geometry content

“Turn-on” Eu(III) chemosensory polymer materials for ammonia and amines detection: Experimental and molecular modeling study

Polymer films doped by Eu(III) complexes are promising candidates for the development of luminescent chemosensors. Here, luminescent properties of Eu(III) carboxylate dibenzoylmethanates in interaction with ammonia and amine vapors have been studied. Quantitative measurements of the optical response showed that with an increase of the analyte concentration in the range of 3–330 ppm, an increase in the luminescence intensity of europium(III) is observed. The reversibility of the luminescent response is established. Quantum chemical calculations showed that ammonia and amine molecules enter not in the Eu(III) coordination sphere, but forms hydrogen bonds between analyte molecules and carbonyl groups of the dibenzoylmethanate ion. Moreover, analyte changes ligand coordination mode: there occur the transformation from tridentate-bridging-cyclic coordination of carboxyl groups to bidentate-bridging which leads to decreasing in the efficiency of cross-relaxation of lanthanide ions. The obtained “turn-on” chemosensory polymer materials are promising for environmental monitoring and food safety testing.

Nanoplasmonic Affinity Analysis System for Molecular Screening Based on Bright‐Field Imaging

AbstractConsidering the improved detection of biological analytes for affinity analysis is highly desirable, a metasurface plasmon resonance (Meta‐SPR)‐based imaging system, incorporating a localized SPR sensing platform with different microfluidic systems and employing simple bright‐field imaging, is established in this study. This system enables low‐level analyte concentration analysis, ranging from 100 pm to 100 nm, with the real‐time removal of nonspecific binding signals within the same device field of view. Combined with microfluidic systems and microdroplet spotting, it is possible to automatically measure the kinetic curves of a sample at ten concentration gradients or detect the specific responses of multiple samples in a single experiment simultaneously. This system can inexpensively and conveniently achieve complex detection functions, demonstrating an innovative breakthrough in sensor detection.

On-Chip Electrochemical Sensing with an Enhanced Detecting Signal Due to Concentration Polarization-Based Analyte Preconcentration.

Here, we integrated two key technologies within a microfluidic system, an electrokinetic preconcentration of analytes by ion Concentration Polarization (CP) and local electrochemical sensors to detect the analytes, which can synergistically act to significantly enhance the detection signal. This synergistic combination, offering both decoupled and coupled operation modes for continuous monitoring, was validated by the intensified fluorescent intensities of CP-preconcentrated analytes and the associated enhanced electrochemical response using differential pulse voltammetry and chronoamperometry. The system performance was evaluated by varying the location of the active electrochemical sensor, target analyte concentrations, and electrolyte concentration using fluorescein molecules as the model analyte and Homovanillic acid (HVA) as the target bioanalyte within both phosphate-buffered saline (PBS) and artificial sweat solution. The combination of on-chip electrochemical sensing with CP-based preconcentration renders this generic approach adaptable to various analytes. This advanced system shows remarkable promise for enhancing biosensing detection in practical applications while bridging the gap between fundamental research and practical implementation.

Internet financial reporting disclosure index of e‐commerce businesses on social media

SummaryThe study measured the Internet Financial Reporting (IFR) disclosure index and compared the results across three continents of the global e‐commerce business. In addition, it documents various social media platforms used by e‐commerce. We use content analysis with a scoring matrix based on content, timeliness, technology, and support used in websites and a one‐way ANOVA. The findings identified an average IFR e‐commerce disclosure index of 0.735, which is of good quality as it approaches the value of 1. There is no difference in index IFR between the three continental zones overall, but slightly different from non‐e‐commerce companies. The results also prove that websites and blog media still dominate the use of social media, while other social media platforms have not provided financial information. Researchers in accounting have not conducted research topics on social media, so there are still limited references and narrow analytical content. This research will interest the e‐commerce business industry and compile their financial reporting through the website to improve the quality of their IFR and financial access. Since the e‐commerce business is an internet‐based company growing significantly, it can use other social media to reveal its reporting as decent work and economic growth. This subject is relatively innovative because none of the IFR disclosure index studies focuses on e‐commerce businesses on social media. It fills the research gap related to the characteristics of e‐commerce businesses, where almost all activities are internet‐based.

Efficient concentration of trace analyte with ordered hotspot construction for a robust and sensitive SERS platform

Surface-enhanced Raman scattering (SERS) platform, which enables trace analyte detection, has important application prospects. By structuring/modifying the surface of the SERS substrate, analyte in highly diluted solutions can be concentrated into localized active areas for highly sensitive detection. However, subject to the difficulty of the fabrication process, it remains challenging to balance hot-spot construction and the concentration capacity of analyte simultaneously. Therefore, preparing SERS substrates with densely ordered hot spots and efficient concentration capacity is of great significance for highly sensitive detection. Herein, we propose an Ag and fluoroalkyl-modified hierarchical armour substrate (Ag/F-HA), which has a double-layer stacking design to combine analyte concentration with hotspot construction. The microarmour structure is fabricated by femtosecond-laser processing to serve as a superhydrophobic and low-adhesive surface to concentrate analyte, while the anodic aluminium oxide (AAO) template creates a nanopillar array serving as dense and ordered hot spots. Under the synergistic action of hot spots and analyte concentration, Ag/F-HA achieves a detection limit down to 10−7 M doxorubicin (DOX) molecules with a RSD of 7.69%. Additionally, Ag/F-HA exhibits excellent robustness to resist external disturbances such as liquid splash or abrasion. Based on our strategy, SERS substrates with directional analyte concentrations are further explored by patterning microcone arrays with defects. This work opens a way to the realistic implementation of SERS in diverse scenarios.

Integrated GIS-based and water quality index for evaluation of groundwater quality in the coastal slum settlements of Lagos, Nigeria

This study aimed to identify drinking water suitability through a statistical and water quality index (WQI) approach in coastal slum settlements in Lagos, Nigeria. A total of 250 borehole groundwater samples collected for drinking purposes during the wet (125) and dry (125) seasons were analysed for their physical and chemical properties from 2019 to 2020. The physical and chemical characterisation of the region was conducted based on the correlation matrix, scree plot, PCA, Q-mode HCA, and ROC curve to identify the types of weathering, the influence of rock on precipitation and evaporation, and types of reactions that affect groundwater composition. The analytical mean concentration results revealed that SO4, Cl, TH, NO3, TDS, and PO4 surpassed the USEPA and WHO permissible limits in both seasons. The WQI results demonstrate that 7.6% and 6.7% of water samples fall into the excellent category, while 12.4% and 11.5% are moderately tolerable. In contrast, for both seasons, >100% of the samples fell into the unsuitable drinking category across SA1-SA5, SB1-SB5, SC1-SC5, SD1−SD5, and SE1−SE5. Groundwater quality mainly deteriorates due to anthropogenic activities and climatic variability. The spatial variation map indicates that groundwater quality is primarily affected during the wet season and, to a lesser extent, in a few slums during the dry season. Seawater influx in coastline areas has been identified as the main factor threatening the quality and availability of groundwater resources. As a result, the groundwater quality is mostly poor to unsuitable, and this study confirms its unsuitability for drinking. Hence, this study serves as a guide for effective action in sustainable groundwater management.

Chemometric-assisted eMIP-modified screen-printed sensor for robust herbicide MCPA determination

The paper describes the development and application of a screen-printed electrode cell with a graphite-ink working electrode modified by a molecularly imprinted electropolymerized polypyrrole for the voltammetric determination of the herbicide 4‑chloro-2-methylphenoxyacetic acid (MCPA). The method exploits the direct measurement of the analyte by applying the differential pulse voltammetry (DPV) technique, taking advantage of the irreversible oxidation peak at about +1.0 V vs. Ag/AgCl pseudo reference electrode. The presence of the molecularly imprinted polypyrrole enhances the sensor's selectivity and sensitivity. A chemometric approach has been crucial for quantitative analysis because of the peak's broad and not well-defined shape. Firstly, a proper pretreatment of the voltammetric signals is identified, proving the most effective is the first-derivative function transformation of the signal. The Partial Least Square regression (PLS) is the tool applied for MCPA quantification. A preliminary PLS model has been developed and validated in dihydrogen phosphate solution at pH 5.5, aiming to optimize the data treatment approach. Then, the same approach is used to develop a PLS model analyzing tap water samples fortified with MCPA and other pesticides as possible interferents to simulate contaminated natural waters. The model correctly predicted the analyte concentration in the range of 2.5–75 μM, assuring the reliability and robustness of the sensor for the possible quantification of MCPA in wastewater samples.

Self-absorption of emission lines in picosecond-laser-produced gold plasmas

Ultrashort laser ablation offers several advantages in various applications compared to traditional nanosecond laser ablation techniques. Despite providing a lower damage threshold, cold ablation with high precision, the impact of self-absorption effects in ultrashort laser-produced plasmas (LPPs) significantly affects the assessment of plasma parameters and analytical outcomes in Laser-Induced Breakdown Spectroscopy (LIBS) analysis. This study investigated the impact of laser energy, analyte concentration, and acquisition gate delay on the self-absorption of emission lines from picosecond LPPs (ps-LPPs) of gold targets at atmospheric pressure. We used four gold targets (24 carat gold, 22 carat gold, 18 carat gold, and Hepatizon) with varying concentrations of gold and copper. To analyze the self-absorption effects in ps-LPP, we examined four neutral emission lines of ps-LIBS spectra (Au I: 267.59 and 627.81 nm; Cu I: 327.98 and 510.55 nm). We observed that with an increase in laser pulse energy, there is a corresponding rise in the self-absorption of emission lines under ambient pressure. This increase in analyte species concentration leads to an elevation in the self-absorption of emission lines. Additionally, as the temporal delay of acquisition extends, self-absorption intensifies.

Acute Inhalation Toxicity Study of 3-Methoxybutyl Acetate in Sprague-Dawley Rats

3-Methoxybutyl acetate is an organic solvent used in a wide range of industrial and consumer applications, such as coatings, adhesives, chemical intermediates, paints, and washing products. We conducted an acute inhalation toxicity study of 3-Methoxybutyl acetate using an inhalation exposure system (nose-only) in Sprague-Dawley rats. Male and female rats were exposed to 3-Methoxybutyl acetate for 4 h/day. The tests were conducted according to the Organisation for Economic Cooperation and Development Guidelines for the Testing of Chemicals (OECD TG) 436. The analytical concentrations of the exposure chamber were 9.76 ± 0.51 mg/L. Clinical signs, mean body weight changes, and gross necropsy results were observed. No deaths, abnormal clinical signs, gross findings at necropsy, or abnormal mean body weight changes related to exposure to the test substance were observed in either sex. Based on these results, we suggested that 3-Methoxybutyl acetate does not have acute toxic effects at the technically acceptable concentration of 10 mg/L.

Nanoplasmonic sensing for studies on liposomes and extracellular vesicles

Nanoplasmonic sensing (NPS) is a comparatively new and powerful technique, which is yet to achieve its full potential across various fields of sciences. The principle behind NPS is localized surface plasmon resonance. NPS has several advantages over traditional sensing techniques when it comes to selectivity, sensitivity, concentration of analyte required, and prohibitive costs involved. The phenomenon exhibits at the resolution depth between two and a few tens of nm, which is difficult in practice or expensive to attain with various modern microscopes. Additionally, NPS is a label free technique which can measure fast and real-time interactions at the solid-liquid interface. This review highlights the applications of NPS in lipid research and the emerging field of extracellular vesicles.

Theory of nanostructured sensors integrated in/on microneedles for diagnostics and therapy

Efficient real-time diagnostics and on-demand drug delivery are essential components in modern healthcare, especially for managing chronic diseases. The lack of a rapid and effective sensing and therapeutic system can result in analyte level deviations, leading to severe complications. Minimally invasive microneedle (MN)-based patches integrating nanostructures (NSs) in their volume or on their surface have emerged as a biocompatible technology for delay-free analyte sensing and therapy. However, a quantitative relationship for the signal response in NS-assisted reactions remains elusive. Existing generalized formalisms are derived for in-vitro applications, raising questions about their direct applicability to in-situ wearable sensors. In this study, we apply the reaction-diffusion theory to establish a generalized physics-guided framework for NS-in-MN platforms in wearable applications. The model relates the signal response to analyte concentration, incorporating geometric, physical, and catalytic platform properties. Approximating the model under NS (binding or catalytic) and environmental (mass transport) limitations, we validate it against numerical simulations and various experimental results from diverse conditions – analyte sensing (glucose, lactic acid, pyocyanin, miRNA, etc.) in artificial and in-vivo environments (humans, mice, pigs, plants, etc.) through electrochemical and optical/colorimetric, enzymatic and non-enzymatic platforms. The results plotted in the scaled response show that (a) NS-limited platforms exhibit a linear dependence, (b) Mass transport-limited platforms saturate to 1, (c) a one-to-one mapping against traditional sensitivity plots unifies the scattered data points reported in literature. The universality of the model provides insightful perspectives for the design and optimization of MN-based sensing technologies, with potential extensions to dissolvable MNs as part of analyte-responsive closed-loop therapeutic applications.

Accurate and reliable surface-enhanced Raman spectroscopy assay for early detection of SARS-CoV-2 RNA with exceptional sensitivity

This research proposes a highly sensitive and simple surface-enhanced Raman spectroscopy (SERS) assay for the detection of SARS-CoV-2 RNA using suitably designed probes specific for RdRp and N viral genes attached to a Raman marker. The sensitivity of the assay was optimized through precise adjustments to the conditions of immobilization and hybridization processes of the target RNA, including modifications to factors such as time and temperature. The assay achieved a remarkable sensitivity down to 58.39 copies/mL, comparable to or lower than the sensitivities reported for commercial fluorescent polymerase chain reaction (PCR) based methods. It has good selectivity in discriminating SARS-CoV-2 RNA against other respiratory viruses, respiratory syncytial virus (RSV), and influenza A virus. The reliability of the assay was validated by testing 24 clinical samples, including 12 positive samples with varying cycle threshold (Ct) values and 12 negative samples previously tested using real-time PCR. The assay consistently predicted true results that were in line with the PCR results for all samples. Furthermore, the assay demonstrated a notable limit of detection (LOD) of Ct (38 for RdRp gene and 37.5 for N-gene), indicating its capability to detect low concentrations of the target analyte and potentially facilitating early detection of the pathogen.

Development of a Reversed-Phase UPLC Method for Assay of Fipronil Including Determination of Its Related Substances in Bulk Batches of Fipronil Drug Substance.

Fipronil is a commonly used pesticide in the agricultural and animal health industries for the protection of crops and control of fleas, ticks, and chewing lice. It is difficult to obtain reproducible retention time and relative retention time (RRT) for a common hydrolytic degradation product of fipronil with the current European Pharmacopeia (EP) monograph for assay and estimation of related substances of fipronil. This situation causes misidentification, mislabeling, and/or false out-of-specification results for this hydrolytic degradation product of fipronil in bulk commercial batches during batch release and/or in the stability samples during the shelf life of a released batch. This study aimed to develop a reversed-phase ultra performance liquid chromatography (UPLC) method for assay and identification of fipronil including identification and estimation of its related substances in bulk drug substance batches of fipronil and provide consistent retention time of the hydrolytic degradation product. Fipronil and its related substances were separated by gradient elution on a Halo C18 column (50 mm × 2.1 mm id, 2.0 µm particle size) maintained at 40°C with 0.1% H3PO4 in H2O as mobile phase-A and acetonitrile-methanol (50 + 50, v/v) as mobile phase-B. Fipronil and its related substances were detected and quantified at 280 nm with a quantitation limit of 0.05% of the target (analytical) concentration. The UPLC method was able to separate all analytes of interest by gradient elution with a total run time of 7 min (approximately 40% faster than EP). In this paper, we report the development and validation of a fast, stability-indicating reversed-phase UPLC method for assay and estimation of related substances of fipronil in stability samples and bulk batches of fipronil. The new UPLC method is approximately 40% faster than the current Ph. Eur. monograph for fipronil assay and the new method provides reproducible retention of a common hydrolytic degradation product of fipronil.

Abstract 3667: Nucleic acid quantification in uveal melanoma aqueous humor

Abstract Introduction: Uveal melanoma (UM) is the most common primary adult intraocular cancer. Though radiation brachytherapy is often employed for UM treatment, half of UM patients develop systemic metastases. In large tumors with macular involvement, the disease and its treatment can lead to blindness. Clinically, UM prognosis is estimated by tumor size, location, American Joint Cancer Commission (AJCC) staging, and histologic findings. Prognostic UM biomarkers have been established but lesions <2 mm in height, particularly near the fovea, are difficult to biopsy, and thus physicians often do not perform biopsies on such tumors. Therefore, the use of aqueous humor (AH) as a liquid biopsy is a promising methodology. The purpose of the study was to quantify analytes in AH samples collected at diagnosis and post-brachytherapy from UM eyes with varying tumor sizes and AJCC Stages to analyze correlations between analyte (dsDNA, microDNA, and protein) concentrations and tumor features. Methods: This case series study included 119 UM AH samples from 66 UM eyes and 16 control AH samples collected from age-matched non-UM glaucoma (GLC) eyes analyzed for unprocessed analytes using Qubit fluorescence assays. Results: Most UM AH samples contained quantifiable analyte concentrations (dsDNA: 94.1%, miRNA: 88.0%, protein: 95.2%) and at higher concentrations than GLC controls (dsDNA, P=0.008; miRNA, P<0.0001; protein, P=0.007). In samples taken at diagnosis, concentrations were higher at more advanced AJCC stages; comparing most advanced Stage III (n=8) to least advanced Stage I (n=24), median dsDNA was 4 times greater (P<0.0001), protein was 3 times greater (P<0.0001), and miRNA was 2 times greater (P=0.001). Analytes were quantifiable in >70% of pre-brachytherapy samples from eyes with tumors <2 mm tall (n=17). Height was positively associated with analyte concentrations at diagnosis (dsDNA: R=0.43, P=0.0007; miRNA: R=0.35, P=0.01; protein: R=0.39, P=0.005). In paired samples (n=53), all analyte concentrations were significantly higher post-therapy compared to diagnosis pre-therapy (Ps<0.01). Conclusions: Although AH analytes were also quantifiable in smaller, less advanced tumors, AH samples from eyes with larger, more advanced tumors had higher analyte concentrations. The detection of prognostic biomarkers in AH may allow for early UM recognition and guide personalized treatment. Citation Format: Christina Chang, Sarah Pike, Mark Reid, Chen-Ching Peng, Benjamin Xu, Jesse Berry, Liya Xu. Nucleic acid quantification in uveal melanoma aqueous humor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3667.

Bioequivalence of zolpidem hemitartrate: new formulations of 5 mg SL, 10 mg and 12.5 mg XR and comparison of their bioavailabilities

Objective: To evaluate pharmaceutical bioequivalence between two formulations of 5 mg zolpidem hemitartrate sublingual and, 10 and 12.5 mg extended-release formulations tablets in healthy subjects under fasting and fed conditions. Methods: An open label, monocentric, randomized, 2 x 2 crossover study in 40 healthy adults under fasting conditions comparing two formulations of zolpidem 5mg sublingual tablets. Analyte concentrations in human plasma were determined using a validated liquid chromatography with a tandem mass spectrometer detector method (LC-MS/MS). The same design was utilized to evaluate the other formulations.
 Results: Statistical analysis has determined geometric mean of test / reference ratio, confidence intervals, and power of the test to the pharmacokinetic parameters Cmax, AUC0-t, and AUC0-∞. The geometric mean ratio (90%CI) of the test drug/reference drug for zolpidem 5mg were 99.89 to 113.57% for Cmax, 97.15% to 108.40% for AUC0-t, and 97.22% to 108.13% for AUC0-∞. Power of the test was 99.35% for Cmax and 100% AUC0-t, and AUC0-∞. Conclusion: Both test and reference are bioequivalent for all formulations and, therefore, they are interchangeable, according to the Brazilian criteria (Anvisa resolution RE nº 1170/2006), since confidence intervals for Cmax and AUC0-t ratios were within 80% and 125%.

Abstract 3636: Myeloid cell rebound associated with GD2-CAR-T cell therapy in solid tumor patients

Abstract GD2 is a validated cancer immunotherapy target with highly restricted normal tissue expression. Promising activity of GD2-CAR-T cell therapy in diffuse midline glioma and neuroblastoma has promoted interest in this therapy. We initiated three phase 1 clinical trials of escalating doses of intravenously administered autologous GD2-CAR-T cells in patients with advanced GD2-positive solid tumors. Emerging evidence suggests that CAR-T cells may increase circulating myeloid derived suppressor cells (MDSCs) and that this event may be mediated and/or triggered by CAR-T cell-derived cytokines and chemokines. These findings may help to explain the limited persistence of CAR-T cell therapy in solid tumor patients. Hence, we sought evidence of MDSC generation and secretion of relevant cytokines and chemokines in our clinical trial patients. The CARPETS trial (www.anzctr.org.au: ACTRN12620000622909) recently closed to recruitment after enrolling 12 patients (7 melanoma, 4 colorectal cancer, and 1 sarcoma). No significant adverse events were attributable to CAR-T cell therapy. We opened two other GD2-CAR-T cell therapy trials in 2023: LEVI’S-CATCH (ACTRN12622000675729) in patients with diffuse midline glioma at Sydney Children’s Hospital and KARPOS (ACTRN12622001514796) in adult patients with recurrent glioblastoma at Royal Adelaide Hospital. To date, 3 patients have enrolled to LEVI’S-CATCH with the first patient clearing the dose limiting (DLT) evaluation period. Three patients completed enrolment to the first dose cohort of KARPOS without DLTs. In all CARPETS patients, CAR-T cell expansion, which was measured by by DNA quantitative PCR and flow cytometry, peaked during the 6-week post-CAR-T infusion DLT evaluation period. In 6 CARPETS patients for whom peripheral blood samples were available, we found by flow cytometry, newly formed or increased size of pre-existing subpopulations of non-classical monocytes and/or MDSCs of mononuclear and/or polymorphonuclear class, which peaked at or after the time of peak CAR-T cell expansion. Typically, serum levels of inflammatory cytokines and chemokines such as TNF, IL-6, IL-8, CCL2, GM-CSF, and CXCL10, were detected only after the CAR-T cell infusion with variable patterns of the timing and concentration of analyte observed among the 12 patients. At the time of writing, blood samples from the first KARPOS patient showed CAR-T cell expansion and the subsequent increases in circulating myeloid derived suppressor cells and inflammatory cytokines. We expect that full data sets for these parameters will be presented for current and future enrolled patients to both studies. To improve CAR-T cell persistence in solid tumor patients, these data indicate the need for new therapeutic strategies to mitigate effects of CAR-T cell induced myeloid cells with antitumor activity. Citation Format: Michael P. Brown, Tessa Gargett, Nga T. Truong, Orazio Vittorio, David S. Ziegler. Myeloid cell rebound associated with GD2-CAR-T cell therapy in solid tumor patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3636.

Abstract 3773: Bladder cancer risk stratification with the Oncuria 10 plex bead-based urinalysis assay using three different Luminex xMAP instrumentation platforms

Abstract Introduction and Objectives: Oncuria® is a bead-based multiplex fluorescence immunoassay that coordinately measures 10 protein biomarkers in urine samples. The current study compared assay performance and output when urine samples were evaluated with the Oncuria assay using three different fluorescence-analyzing instruments commonly used in diagnostic laboratories worldwide. Methods: We compared the performance of the clinically validated Oncuria bladder cancer (BC) multiplex immunoassay when data output was generated on three different analyzer systems. Voided urine samples from 36 subjects (18 with BC and 18 Controls) were reacted with Oncuria test reagents in three 96 well microtiter plates, and consecutively evaluated on the LED/image based MagPix, and laser/flow based Luminex 200 and FlexMap 3D (all xMAP instruments from Luminex Corp., Austin, TX). The BC assay uses magnetic bead based fluorescence technology (xMAP, Multi-analyte profiling; Luminex) to simultaneously quantify 10 protein analytes in urine specimens [i.e., angiogenin (ANG), apolipoprotein E (ApoE), carbonic anhydrase IX (CA9), CXCL8/interleukin-8 (IL-8), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase-10 (MMP-10), serpin A1/alpha-1 anti-trypsin (A1AT), serpin E1/plasminogen activator inhibitor-1 (PAI-1), CD138/syndecan-1 (SDC1), and vascular endothelial growth factor-A (VEGF-A)]. Results: All three platforms categorized all 10 analytes in identical samples at nearly identical concentrations, with variance across systems typically <5%. While the most contemporary instrument, the FlexMap 3D, output higher raw fluorescence values than the two comparator systems, standard curve slopes and analyte concentrations determined in urine samples were concordant across all three units. Forty-four percent of BC samples registered ≥1 analyte above the highest standard concentration, i.e., A1AT (n=7/18), IL-8 (n=5), and/or ANG (n=2). In Controls, A1AT was higher in one sample. Conclusions: In conclusion, the Oncuria BC assay performed similarly well across three different flow analysis platforms for all 10 analytes simultaneously evaluated in urine samples. This agreement across instruments indicates that the test is amenable to standardized performance in laboratories using existing xMAP, without requiring costly outlays for new equipment. Citation Format: Sunao Tanaka, Toru Sakatani, Kaoru Murakami, Richard T. Waldron, Walter Morales, Wayne Hogrefe, Charles J. Rosser, Hideki Furuya. Bladder cancer risk stratification with the Oncuria 10 plex bead-based urinalysis assay using three different Luminex xMAP instrumentation platforms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3773.