OncoDiscover Blood Test Detects Cancer Relapse Early | Dr. Jayant Khandare Expert Insights | August 28, 2022 Revolutionary Blood Test Detects Cancer Relapse Early | Dr. Jayant Khandare Discover India's first approved, highly affordable blood test that detects cancer relapse and metastasis earlier than traditional CT or MRI scans. Developed by Dr. Jayant Khandare and clinically validated with Tata Memorial Hospital, this painless test catches circulating tumor cells before it's too late. Cancer relapse and metastasis are responsible for the vast majority of cancer-related fatalities, often because traditional imaging tools like CT and MRI scans fail to catch the disease's return in time. This video highlights a groundbreaking and highly affordable blood test—India's first-in-class medical device approved by the Drug Controller General of India. Developed by Dr. Jayant Khandare, the OncoDiscover test requires only a small volume of blood to identify circulating tumor cells (CTCs) and detect minimal residual disease before it becomes uncontrollable. Clinically validated in partnership with Tata Memorial Hospital in Mumbai, this painless innovation provides oncologists with a powerful tool to monitor patients, optimize treatment, and ensure families never have to hear the dreaded words, "It's too late." Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Preoperative CTC levels predict prognosis and survival in oral squamous cell carcinoma. Publications | July 1, 2022 Manuscript: Circulating tumor cells as a predictor for poor prognostic factors and overall survival in treatment nay¨ve oral squamous cell carcinoma patients Preoperative circulating tumor cell levels strongly correlate with metastasis, disease severity, and reduced survival in oral squamous cell carcinoma patients. Objective: The aim of this study was to investigate the presence of circulating tumor cells (CTCs) and their correlation with prognostic factors and clinical outcomes in treatment-naive patients with oral squamous cell carcinoma. Study design: CTCs were isolated using the OncoDiscover technique from presurgically obtained peripheral blood of 152 patients with treatment-naive oral squamous cell carcinoma. Sensitivity analysis was performed by including 40 healthy controls. CTC cutoff values for clinicopathologic factors were obtained from receiver operating characteristic curves. Multivariate models determined the significance of CTCs as independent variables. Kaplan–Meier analysis differentiated overall survival based on CTC values corresponding to disease stage. Results: Sensitivity, specificity, and accuracy of CTC detection were 94.32%, 98%, and 95.17%, respectively. The platform differentiated true positives at >3.5 CTCs (P < .00001). CTC counts above 20.5 were suggestive of nodal metastasis (P < .0001), with a linear trend for detecting occult metastasis (P = .061). Early and advanced stages could be differentiated by >13.5 CTCs (P < .0001). Elevated CTC levels were significantly associated with extranodal extension (>21.45 CTCs, P = .025), perineural invasion (>19.35 CTCs, P = .049), and depth of invasion (>12.5 CTCs, P = .0038). Median survival was reduced by 19 months when CTC levels were >13. Conclusions: Preoperative CTC levels demonstrated a strong correlation with adverse clinicopathologic factors and suggested their role as a sensitive prognostic marker for predicting survival outcomes and disease progression. (Oral Surg Oral Med Oral Pathol Oral Radiol 2022;134:73–83) View Manuscript Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

CTC-based PD-L1 and EGFR detection supports targeted therapy in lung cancer. Publications | October 24, 2023 ESMO 2023: Expression of PD-L1 and EGFR on circulating tumor cells in advanced Lung cancer patients CTC analysis using OncoDiscover® enables dynamic detection of PD-L1 and EGFR targets in advanced lung cancer, supporting personalized targeted and immunotherapy decisions. Background: Targeted molecular therapy and immunotherapy have revolutionized the treatment of advanced lung cancer (ALC). Although therapeutically significant, the outcome of immune checkpoint inhibitors (ICI) or tyrosine kinase inhibitors (TKI) depends on the presence of their respective targets in tumor cells. Evaluating targets based on solid tissue biopsy may often be misleading, particularly in progressive patients despite therapy administration. Additionally, tissue biopsy provides a static signature of target protein expression from an evolving tumor. The unmet need for dynamic detection and monitoring of actionable targets could be addressed by circulating tumor cells (CTCs). Here, we report on the utility of CTCs to detect actionable targets in advanced lung cancer (ALC) patients. Methods: We retrospectively analyzed 193 ALC patients for programmed death-ligand 1 (PD-L1) and EGFR expression on CTCs. CTCs were isolated using the Drug Controller General of India-approved OncoDiscover technology based on immunomagnetic targeting using anti-EpCAM antibodies and immunostaining with anti-EGFR and PD-L1 antibodies. CTCs were detected based on the expression of cytokeratins (CKs), absence of CD45, and prominent DAPI-stained nuclei. The presence or absence of EGFR and PD-L1 was determined using automated immunofluorescence microscopy. Results: Among the evaluated cohort, 67% of patients showed the presence of CTCs with a mean value of 4.2 (range: 1 to 62; SD = 10.65). The absence of CTCs in the remaining 33% of patients could be attributed to therapy response in clinically stable disease. Among all patients showing the presence of CTCs, 66% showed detectable expression of PD-L1, while 42% showed strong expression of EGFR. The presence of PD-L1 demonstrated a significant association with CTCs. Similarly, the expression of EGFR among detected CTCs showed high significance compared to reported tissue biopsy data in the literature. Conclusions: Detection of therapeutic targets on CTCs obtained from advanced lung cancer patients strongly indicates that these patients may qualify for anti-EGFR and PD-L1 targeted therapies. Systematic studies with larger sample sizes are required to further strengthen liquid biopsy–based detection of actionable targets. This approach could significantly benefit advanced lung cancer patients showing progressive disease despite chemotherapy or radiotherapy. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Dual ctDNA and CTC biomarkers improve detection of cancer progression. Publications | June 3, 2024 ASCO 2024: Impact of ctDNA genomic mutations and CTCs biomarker duo on clinical concordance in localized, progressive, and metastatic disease. Dual biomarker analysis of ctDNA and circulating tumor cells reveals disease progression and metastasis across multiple cancer types. Background Genomic mutations identified from circulating tumor DNA (ctDNA) have been shown to correlate positively with clinical disease status. EGFR and intracellular cell progression and proliferation pathways involving BRCA1/2 and TP53 genes drive high ctDNA load in progressive cancer patients. Circulating tumor cells (CTCs) indicate cellular residual disease (CRD). Together, ctDNA and CTCs as dual biomarkers offer predictive insights into tumor progression and metastasis, which may be valuable for early detection and treatment modifications. Methods In a retrospective study, 96 cancer patients (including lung, colorectal, breast, stomach, and other cancers) who had recently undergone treatment were investigated for the presence of CTCs and genomic mutations from ctDNA using the OncoMonitor test. Libraries were prepared using a hybridization-capture method covering 1000 targets with a mean sequencing depth of 5000× on the Illumina NextSeq 2000 platform. The test detected genomic alterations including single nucleotide variations (SNVs), small insertions and deletions (INDELs), copy number variations (CNVs), and translocations (fusions) using a 96-gene panel. CTCs were isolated using the OncoDiscover platform and identified as CK18+, PD-L1+, CD45- cells in 1.5 ml of blood. Results Among the 96 pan-cancer patients, 15.6% (n = 15) were identified with localized progressive disease without metastasis based on radiological findings, of which 60% (n = 9) showed at least one genomic alteration detected from ctDNA. Additionally, 12.5% (n = 12) patients were identified with metastatic disease from radiological findings, of which 58.3% (n = 7) showed the presence of at least one CTC. Among these, 33.3% (n = 4) patients had two CTCs, while five patients had no detectable CTCs. Furthermore, metastatic patients showed ctDNA load in 66.6% (n = 8) of cases with at least one genomic finding. In the metastatic disease cohort, CTC enumeration showed a concordance of 58.3% (n = 7) with metastatic radiological findings, while genomic findings from ctDNA showed a concordance of 66.6% (n = 8) with metastatic radiological findings. Among 23.9% (n = 23) patients identified radiologically with stable or treatment-responsive disease, 73.9% (n = 17) had no detectable genomic mutations from ctDNA, and 26.0% (n = 6) were CTC-negative, consistent with radiological findings. However, 26.0% (n = 6) patients had at least one genomic finding, contributing to discordance with radiological findings. Overall, genomic findings from the dual biomarkers showed concordance with radiological findings in 26.6% (n = 4) patients with progressive disease, 41.6% (n = 5) patients with metastatic disease, and 17.3% (n = 4) patients with stable or treatment-responsive disease. Conclusions Patients with progressive and metastatic disease identified through radiological findings showed concordance with dual ctDNA and CTC biomarkers. The concordance of ctDNA in progressive disease and CTC detection in metastatic disease highlights the individual significance of these biomarkers and supports their combined use for monitoring disease status and guiding treatment decisions. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Antibody-coated cotton substrate for CTC enumeration and chemotherapy response. Publications | February 18, 2022 Manuscript: Antibody mediated cotton-archetypal substrate for enumeration of circulating tumor cells and chemotherapy outcome in 3D tumors Cotton microfluidic substrate enables efficient CTC isolation, 3D tumor growth, and drug response testing for improved cancer diagnostics and therapy research. Circulating tumor cells (CTCs) are distinct cancer biomarkers established in clinical settings for early cancer detection, metastasis progression, and minimal residual disease (MRD) monitoring. Despite numerous advances, comprehensive molecular characterization of CTCs remains extremely challenging due to their rarity and heterogeneity. Here, we present a novel cotton microfluidic substrate (CMS) as an innovative biomedical matrix that efficiently isolates CTCs while facilitating in vitro CTC expansion, enabling further downstream analysis of these rare cells. CMS enabled both static and dynamic isolation of cells from the MCF-7 cancer cell line, as well as from the blood of head and neck squamous cell carcinoma (HNSCC) patients. The cell capture efficiencies were further compared with the clinically regulated OncoDiscover® Liquid Biopsy Test. Furthermore, CMS served as a matrix on which the captured cancer cells were grown into 3D tumor models to study anti-cancer drug efficacy and multi-drug resistance (MDR) mechanisms. The design of the CMS employed two different surface chemistries—flattened and nanostructured surfaces—each conjugated with anti-EpCAM antibodies to evaluate CTC capture efficiency and 3D tumor growth dynamics. The nanostructured surface was highly efficient in capturing CTCs and promoted 3D tumor spheroid formation, showing a five-fold increase in size from day 3 to day 10 of culture. Moreover, when treated with the anti-cancer drug cisplatin, an almost half reduction in tumor size was achieved within 24 hours, followed by a cytostatic threshold and the eventual acquisition of drug resistance within three days. Conclusively, the CMS matrix exhibits potential for the further development of “tissue-on-chip” and “point-of-care” medical devices in cancer diagnostics, as well as for evaluating chemotherapeutic efficacy in drug discovery and development. View Manuscript Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Chemo-specific designs for the enumeration of CTCs: advances in liquid biopsy Publications | December 18, 2020 Manuscript: Chemo-specific designs for the enumeration of circulating tumor cells: advances in liquid biopsy Review on chemo-specific nano/micro substrates for efficient CTC isolation, enabling liquid biopsy, metastasis detection, and real-time cancer monitoring. Advanced materials and chemo-specific designs at the nano- and micrometer scale have ensured revolutionary progress in next-generation clinically relevant technologies. For example, isolating a rare population of cells, such as circulating tumor cells (CTCs) from blood among billions of other blood cells, is one of the most complex scientific challenges in cancer diagnostics. Achieving this level of exceptional specificity for extracellular biomarker interactions requires chemical tunability through the use of advanced materials and multistep reactions in both solution and insoluble states. This review delineates the chemo-specific substrates, chemical methods, and structure–activity relationships (SARs) of chemical platforms used for the isolation and enumeration of CTCs, thereby advancing the relevance of liquid biopsy in cancer diagnostics and disease management. We highlight the synthesis of cell-specific, tumor biomarker-based chemo-specific substrates utilizing functionalized linkers through chemistry-based conjugation strategies. The capacity of these nano- and micro-scale substrates to enhance interaction specificity and efficiency with targeted tumor cells is discussed in detail. Furthermore, this review emphasizes the importance of CTC capture and downstream processes involving genotypic and phenotypic CTC analysis in real time. These approaches enable early detection of metastasis progression, evaluation of chemotherapy treatment response, and monitoring of progression-free survival (PFS), disease-free survival (DFS), and overall survival (OS) in cancer patients. Royal Society of chemistry. View Manuscript Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

AI-based image analysis rapidly profiles CTC morphology and biomarker expression. Publications | September 16, 2025 Profiling of PD-L1 and HER2 over expression on cancer cells using AI based macro-driven automation AI-based image analysis rapidly profiles circulating tumor cells, quantifying morphology and biomarkers like PD-L1 and HER2 for cancer research. Abstract Background Extravasation, invasion, epithelial-to-mesenchymal transitions, metastasis progression, immune evasion, and therapeutic resistance are driven by phenotypic alterations in cancer cells. Assessing cell morphology, stiffness, and deformability is therefore crucial. The expression of PD-L1, HER2, EGFR, and cytokeratins (CKs) serves as key phenotypic biomarkers for precision oncology. We developed an AI-based image analysis tool that rapidly captures these transitions in cell assays, including specific protein biomarkers expressed on circulating tumor cells (CTCs). Methods We extended an ImageJ macro to enable rapid and reproducible extraction of biophysical parameters. The macro processes .lif, .nd2, and .czi file formats, using DAPI for nuclear segmentation and fluorophore-conjugated antibodies to delineate cytoplasmic boundaries. We evaluated automatic channel detection, intensity normalization, Otsu thresholding, and per-cell quantification of parameters such as surface area, circularity index (CI), and mean fluorescence intensity. Violin plots illustrated temporal variations in CI across A549 and MCF7 cells. Validation was conducted on CTCs isolated from cancer patient samples (n = 100) for PD-L1 and HER2 expression. Results The macro reduced image processing time from 7 minutes to 3 seconds per sample. A549 cells showed higher and more consistent CI values across all time points, while MCF7 cells demonstrated lower CI with greater variability, particularly at 24 and 72 hours. Quantitative measurements of PD-L1 and HER2 expression showed 100% concordance between the ImageJ macro and Zeiss software outputs, confirming analytical accuracy. CK18 intensity (~60–400) and PD-L1 (~20–50) levels measured by both platforms validated the macro’s ability to detect a wide range of marker expression in CTC subsets. CTCs exhibited higher CI values and greater morphological heterogeneity, consistent with their invasive phenotype. Conclusions We present an AI-driven macro that quantifies the biophysical characteristics of cancer cells, enabling precise phenotypic profiling, including circularity index, proliferation rates, and overexpression of biomarkers such as PD-L1 and HER2 in both cultured cell lines and patient-derived CTCs. View Publication Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

AACR 2026 - Actorius Innovations and Research Events | April 23, 2026 AACR 2026 Highlights | April 17-22 | San Diego, California Highlights from AACR 2026 Highlights from AACR 2026 We exhibited and presented two scientific posters that highlight the evolving role of circulating tumor cells (CTCs) in cancer progression and management: Over expressing PD-L1 circulating tumor cells with clusters in prostate cancer patients. Depletion of circulating tumor cells using an automated device with non-hemolytic affinity-based substrates These studies reflect our continued efforts to better understand CTC biology—not only as indicators, but as active contributors to metastasis. At the center of this work is OncoMetastat® , our patented investigational extracorporeal blood-processing platform. It is envisaged to selectively capture and study circulating tumor cells from a patient’s bloodstream while maintaining blood integrity. Designed with a strong focus on precision and safety, the current prototype is under evaluation for its potential relevance in metastasis research and broader cancer management. Some Interesting Clicks Video Highlights We look forward to engaging with researchers, clinicians, and innovators shaping the future of oncology. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Magnetic nanocrystals capture tumour cells from blood samples Press Release | April 11, 2024 Magnetic nanocrystals capture tumour cells from blood samples These nanomaterials could speed up discovery of anti-cancer drugs Cellulose-based magnetic nanocrystals and nanofibres can capture circulating tumour cells (CTCs) from the blood samples of head and neck cancer patients. A magnet is used to separate the trapped tumour cells, which are then identified under a fluorescence microscope. This technique could potentially be used to monitor cancer progression in real time, says an international team, which included researchers at North Dakota State University, USA, the Tata Memorial Hospital in Mumbai, and Actorius Innovations and Research, and Dr. Vishwanath Karad MIT World Peace University, both in Pune. Click the below link to read the full article. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

OncoDiscover detects CTCs, PD-L1 and clusters for longitudinal TNBC monitoring and care. Publications | August 30, 2026 Evaluation of circulating tumor cells expressing PD-L1 and CTC clusters at baseline and follow ups in triple-negative breast cancer A retrospective study of 180 TNBC blood samples evaluated CTC burden, PD-L1 expression and CTC clusters using the CDSCO-approved OncoDiscover platform, highlighting their potential for longitudinal disease monitoring and biomarker-guided management. Introduction: Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype characterized by a high risk of recurrence and metastasis. Assessment of circulating tumor cells (CTCs), along with evaluation of PD-L1 expression and CTC clusters, provides prognostic, minimal cellular residual disease (MCRD), and predictive information. The duration of longitudinal CTC monitoring has not been established. This study evaluates the implications of detecting CTC burden, PD-L1 expression, and CTC clusters in TNBC patients at baseline and during follow-up for improved clinical outcomes. Methods: This retrospective observational study analyzed 180 peripheral blood samples from patients with TNBC, including 141 baseline (BL) and 39 longitudinal follow-up (FU) samples. Peripheral whole blood (1.5 mL) was processed using the CDSCO-approved OncoDiscover liquid biopsy platform, which employs antibody-mediated immunomagnetic enrichment and automated fluorescence microscopy to identify EpCAM⁺/CK18⁺/DAPI⁺/CD45⁻ CTCs. CTC positivity was defined as ≥1 confirmed CTC. Descriptive statistics were used to evaluate CTC burden, PD-L1-positive CTCs, and CTC clusters across BL and FU cohorts and age groups. Results: CTCs were detected in 114/180 (63.3%) patients, while 66 (36.7%) were CTC-negative. PD-L1-positive CTCs were identified in 62/114 (54.4%) CTC-positive patients (34.4% of the overall cohort), and CTC clusters were detected in 17/180 (9.4%) patients. The overall mean CTC count was 0.91, with higher values observed in FU than BL samples (1.18 vs. 0.81). Mean CTC cluster counts were also marginally higher in FU samples (0.10 vs. 0.09). Most CTC-positive patients harbored a single CTC (77/180, 42.8%), followed by two (14.4%), three (5.0%), and four (1.1%) CTCs. Patients aged 20–30 years demonstrated the highest mean CTC count (1.83), whereas those aged 71–80 years had the lowest mean CTC count (0.50). Conclusion: The OncoDiscover platform detected CTCs in nearly two-thirds of patients with TNBC and demonstrated that more than half of CTC-positive patients expressed PD-L1. Simultaneous assessment of CTC enumeration, PD-L1 expression, and CTC clusters represents a promising minimally invasive strategy for longitudinal disease monitoring and biomarker-guided management in TNBC. Monitoring changes in CTCs and the duration of longitudinal MCRD from baseline remains critical, although this is not yet well established. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

OncoMetastat safely captured cancer cells across multiple onco-dialysis cycles in rats. Publications | August 30, 2026 Capture of Cancerous Cells and Overall Survival in Rats using Ex-vivo Extracorporeal Device in Multiple Onco-dialysis Cycles An animal study evaluated OncoMetastat for ex-vivo cancer cell capture, safety, and survival in rats. Across multiple onco-dialysis cycles, cancer cells were captured, all test-group rats survived 14 days, and histopathology showed normal tissues. Introduction: The micrometastatic cascade involves the progression of cancer cells from the primary tumor to distant organs. Capture and elimination of circulating tumor cells (CTCs) may potentially delay metastasis; however, demonstrating this in ex-vivo models remains highly challenging. Here, we demonstrate cancer cell capture, safety, and overall survival in rats using an ex-vivo extracorporeal device across multiple onco-dialysis cycles. Methods: In this animal observational study (IAEC/2025-26/M2/15), 12 White Wistar Albino rats were subjected to onco-dialysis using the miniature OncoMetastat device containing anti-EpCAM antibody-coated glass beads. For safety evaluation, rats were anesthetized, and 1 ml of blood was obtained from the retro-orbital artery, heparinized, and exposed to the device. Subsequently, 0.3 ml of blood was re-injected through the tail vein, and the animals were monitored for changes in temperature and signs of anaphylactic shock. DAPI-pre-stained MCF-7 cells (0.5 × 10⁶) were injected via the rat tail vein (n=6). After 15 minutes, 1 ml of blood was withdrawn from the retro-orbital artery, heparinized, circulated through the OncoMetastat device, and 0.3 ml was recirculated into the rats. Glass beads were analyzed for cancer cell capture across cycles I–III, with 48 hours of rest between cycles. Animals were monitored for 14 days for overall survival (OS) and subsequently sacrificed for histopathological evaluation. Results: Eleven of the 12 rats demonstrated safety following exposure to the onco-dialysis device. A total of 23 cells were captured across three rats over three cycles. In Rat 1, 9, 5, and 1 cells were captured in cycles I, II, and III, respectively, with a mean of 5.0 cells. In Rat 2, 3, 2, and 1 cells were captured across cycles I–III, respectively. In Rat 3, 1, 1, and 0 cells were captured across cycles I–III, respectively. The mean distribution of captured cells across all rats was 7.67/ml. Overall survival was observed in both groups, although one rat in the control group experienced mortality. All rats in the test group survived the 14-day observation period. Histopathological evaluation revealed normal tissues in device-exposed rats. Conclusion: This study demonstrates the safety and capture of cancer cells using an ex-vivo extracorporeal device across multiple onco-dialysis cycles in rats, with all test-group animals surviving the 14-day observation period and showing normal tissue histopathology. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

CK18 variance in CTCs across cancer types necessitates validated enumeration. Publications | October 9, 2021 ESMO 2021: Validation of Cytokeratin (CK18) Protein Expression in Epithelial Cell lines and in Circulating Tumor Cells (CTCs) Study shows significant CK18 expression variance across different cancer cell lines and CTCs, highlighting the need for regulated enumeration tools. Background CTCs predict an unfavourable prognosis and outcomes in cancers. Lowering of cytokeratin 18 expression is a hallmark of epithelial mesenchymal transition (EMT). Homogeneity and validation of CK18 expression in cancer cell lines and CTCs originating from distinct solid tumors is indeterminate and may contribute to non-specific counts. We hypothesize that the expression of CK18 in varied cell lines may differ quantitatively, and additionally may exhibit similar trends in CTCs enumerated from different tumor types. Methods CK18 variance in epithelial cell lines (e.g., A549+, MCF-7+, and MEF-) (n=192,269 cells) and CTCs (n=63) of different phenotypes was analyzed and compared. The fluorescence intensity was measured post-immunostaining, using motorized-automated, computer-assisted scanning, and through a customized ImageJ macro tool. The effect of anti-CK18 concentrations (0.06-6 μg/ml) and binding constants (Kb) was measured across all cell lines. CTCs were enumerated from head and neck squamous cell carcinoma (HNSCC) patients' blood samples (CTRI/2018/03/012905) and from clinical samples (e.g., breast, lung, colorectal (CRC), ovarian) using the clinically relevant OncoDiscover platform. Results CK18 mapping revealed diverse fluorescence intensities distribution in three cell lines, as well as in HNSCC, lung, breast, ovarian, and colorectal cancer CTCs (Table). In addition, the protein binding assay showed 8.65 x 10^3 Kb (M^-1) for MCF7 and 7.9 x 10^3 for A549 cells indicating concentration-dependent binding for CK18 expressing proteins on cells and may be varied in CTCs of different cancer types. Compared to the CK- cell line (MEF), the normalized CK18 intensity was higher by 290% and 310%, respectively, in MCF7 (breast) and A549 (lung) cells, demonstrating the variation in CK18 expression. On the other hand, CTCs showed significant diversity in CK18 expression with buccal mucosa revealing the lowest (67%), while CTCs of CRC origin demonstrated the highest expression (320%) (Table). CK18 intensity was represented across the cell lines and on CTCs enumerated from different cancer types. Conclusions Non-regulated CTC enumeration platforms pre-requisite critical validations to eliminate the non-specificity of CTC counts, which are highly imperative to clinical decisions in cancer management. Clinical Trial Identification CTRI/2018/03/012905. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe