Agency: European Commission | Branch: FP7 | Program: CP | Phase: ICT-2009.3.9 | Award Amount: 9.20M | Year: 2010
Cancer remains a prominent health concern afflicting modern societies. Continuous innovations and introduction of new technologies are essential to level or even reduce current healthcare spending. As the analysis of occult tumour cells (OTC) in blood or bone marrow is most promising in this respect, MIRACLE aims to develop a low-cost, fully automated, integrated lab-on-a-chip (LOC) system for the isolation, counting and characterization of OTCs starting from clinical samples.A major challenge for OTC detection is their extremely low concentration (below a single cell per mL) in clinical samples. Current detection methods are often based on enrichment techniques followed by cumbersome microscopic analysis of the cell phenotype. Some of these procedure steps have been semi-automated (Cellsearch, J&J), but the interpretation of the cell morphology requires expertise and remains partially subjective. In contrary to standard phenotyping tests, MIRACLE aims to determine the genotype by integrating all sample processing and detection steps in a miniaturized system. This envisaged, fully-automated MIRACLE test would yield decisive results within half a day for less than 50 EUR, as compared with contemporary diagnostics tests that may take days.With the essential individual modules recently demonstrated on automated chips in a joint project by some of the partners involved (MASCOT FP6 027652), the MIRACLEs consortium is uniquely positioned to lead the projects main objectives to a successful outcome, well ahead of the current state-of-the-art. Combining the teams multidisciplinary and unique expertise avoids unnecessary overlap. Integrating all components into a fully operational LOC platform will represent an immense advance for Europe to cope with interfacing and integration problems generic to microfluidic and smart miniaturized systems. More importantly, the realisation of the MIRACLE vision will revolutionise cancer diagnostics and individualized theranostics.
Kaprio T.,University of Helsinki |
Hagstrom J.,University of Helsinki |
Fermer C.,Fujirebio Diagnostics AB |
Mustonen H.,University of Helsinki |
And 4 more authors.
BMC Cancer | Year: 2014
Background: Podocalyxin (PODXL) is a transmembrane sialomucin, whose aberrant expression and/or allelic variation associates with poor prognosis and unfavourable clinicopathological characteristics in different cancers. Membranous expression of PODXL has been suggested to be an independent marker of poor prognosis in colorectal cancer (CRC), and previously by an in-house monoclonal antibody, we showed that also cytoplasmic overexpression of PODXL predicts poor prognosis. The aim of this study was to compare two PODXL antibodies with different epitopes case-by-case in CRC patients.Methods: Of 840 consecutively operated CRC patients from Helsinki University Central Hospital, PODXL expression by polyclonal HPA 2110 antibody was evaluated from 780. Associations of PODXL expression with clinicopathological parameters and the impact of PODXL expression on survival were assessed. Kappa-value was used to assess the comparability of the two antibodies.Results: Membranous PODXL expression associated with unfavourable clinicopathological parameters and with higher risk for disease-specific death from CRC within 5 years (unadjusted hazard ratio (HR) = 1.90; 95% confidence interval (CI) (1.32-2.75); adjusted HR = 1.64; 95% CI (1.11-2.43)). The comparability of expressions by the two antibodies was low (kappa =0.219, standard error 0.060, p < 0.0001). Combination of two antibodies identified a group of patients with even worse prognosis (unadjusted HR = 6.00; 95% CI (3.27-13.0); adjusted HR = 2.14; 95% CI (1.12-4.07)).Conclusion: Membranous expression by the polyclonal PODXL antibody and cytoplasmic overexpression by the monocolonal PODXL antibody are both independent markers of poor prognosis, but they recognise different groups of patients, both of which have poor prognosis. The combined use of the antibodies reveals a group with an even worse prognosis. The biological reasons for the difference between antibodies warrant further studies. © 2014 Kaprio et al.
PubMed | University of Hamburg, Innsbruck Medical University, Charité - Medical University of Berlin, International Health Sciences University and 3 more.
Type: Journal Article | Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine | Year: 2016
Patients with epithelial ovarian cancer (EOC) are at high risk of tumor recurrence. Human epididymis protein 4 (HE4) has been shown to be overexpressed in EOC. The primary aim of our study was to evaluate the role of HE4 in predicting recurrence in EOC patients. Furthermore, we assessed the role of HE4 in predicting recurrence after second-line chemotherapy. We retrospectively analyzed data of 92 out of 275 primary EOC patients of the multicenter project Ovarian Cancer: Diagnosis of a silent killer (OVCAD). The concentrations of HE4 and CA125 were determined preoperatively and 6months after the end of platinum-based first-line chemotherapy (FU) using ELISA and Luminex technique, respectively. The role of HE4 and CA125 for prediction of recurrence was determined using receiver operating characteristics (ROC) curves. Out of 92 patients included, 70 (76%) were responders and 22 (23%) non-responders in terms of response to platinum-based first-line chemotherapy. Median HE4 concentrations at follow-up (FU) differed between responders and non-responders (60.5 vs. 237.25pM, p=0.0001), respectively. The combined use of HE4 and CA125 at FU with cut-off values of 49.5pM and 25U/ml for HE4 and CA125, respectively, for predicting recurrence within 12months after first-line chemotherapy performed better than HE4 or CA125 alone (area under the curve (AUC) 0.928, 95% confidence intervals (CI) 0.838-1, p<0.001). HE4 at FU could predict recurrence within 6months after second-line chemotherapy (AUC 0.719, 95% CI 0.553-0.885, p=0.024). The combination of both elevated biomarkers revealed significantly worse estimated median progression-free survival (PFS; hazard ratio (HR) 8.14, 95% CI 3.75-17.68, p<0.001) and slightly worse PFS in those in whom only one biomarker was elevated (HR 1.46, 95% CI 0.72-2.96, p=0.292) compared to those patients in whom no biomarker was elevated. For the estimated median overall survival (OS), our analysis revealed similar results. HE4 in combination with CA125 performed better than CA125 and HE4 alone in predicting recurrence within 12months after first-line chemotherapy.
Agency: European Commission | Branch: FP7 | Program: CP-TP | Phase: HEALTH-2007-1.1-4 | Award Amount: 3.95M | Year: 2008
The PROACTIVE consortium will through research and innovation develop the multiplexed proximity ligation assay into a high throughput protein detection and quantification technology along with novel data management and analysis tools. Hundreds of putative biomarkers of the plasma proteome can then be assayed with high sensitivity in minute sample volumes, far surpassing any current capabilities. This consortium combines three SMEs across Europe with synergistic competences in technology development, reagent manufacturing, and software development for data management and analysis. Together with clinical cancer scientists, biostatisticians, and diagnostics industry, these high capacity tools will be evaluated in pilot projects for cancer biomarkers using biobanked samples. Preliminary data in the literature claim improved diagnostics with the use of multiple complementary protein markers. However, there is a lack of suitable high throughput procedures for finding new markers and discovering which markers complement each other into effective diagnostic panels. Better capabilities to diagnose cancer at the early and most curable stages will greatly improve human health and reduce health care costs. Patient stratification is also in need of better diagnostics to facilitate the selection of appropriate patient care. Many clinically used immunoassays capable of diagnosing cancer have been in use for many years as single markers but with limited sensitivity and specificity. No marker can today single-handedly diagnose all cases with desired accuracy for a certain cancer type. Also, the performance of these markers is decreased for the earliest stages of the disease. At the conclusion of this collaborative project, the research and innovation conducted within the consortium will enable the partners to position themselves at the very fore-front of high-throughput biomarker research strengthening their competitiveness in the international arena.
Laborla N.,Rovira i Virgili University |
Fragoso A.,Rovira i Virgili University |
Kemmner W.,Charité - Medical University of Berlin |
Latta D.,Fraunhofer Institute of Microtechnology Mainz |
And 5 more authors.
Analytical Chemistry | Year: 2010
Detection of proteins that signal the presence or recurrence of cancer is a powerful therapeutic tool for effective early diagnosis and treatment. Carcinoembryonic antigen (CEA) has been extensively studied as a tumor marker in clinical diagnosis. We report on the development of an amperometric biosensor for the detection of CEA based on the immobilization of anti-CEA monoclonal antibody on a novel class of bipodal thiolated self-assembled monolayers containing reactive N-hydroxysuccinimide (NHS) ester end groups. The current variations showed a linear relationship with the concentration of CEA over the range of 0-200 ng/mL with a sensitivity of 3.8 nA·-mL·ng -1 and a detection limit of 0.2 ng/mL, which is well below the commonly accepted concentration threshold (5 ng/mL) used in clinical diagnosis. Real time and accelerated stability studies of the reporter antibody under various storage conditions demonstrated that the enzymatic activity and antibody affinity of the conjugate is retained for long periods of time in commercial stabilizing buffers such as StabilGuard Biomolecule Stabilizer, and a prediction of the stability trends was carried out using the kinetic and thermodynamic parameters obtained from the Arrhenius equation. The developed immunosensor as well as a commercially available enzyme-linked immunosorbent assay (ELISA) kit were successfully applied to the detection of CEA in serum samples obtained from colon cancer patients, and an excellent correlation of the levels of CEA measured was obtained. Ongoing work is looking at the incorporation of the developed biosensor into a platform for multiplexed simultaneous detection of several breast cancer related biomarkers © 2010 American Chemical Society.
Braicu E.I.,Charité - Medical University of Berlin |
Fotopoulou C.,Charité - Medical University of Berlin |
Van Gorp T.,Catholic University of Leuven |
Richter R.,Charité - Medical University of Berlin |
And 11 more authors.
Gynecologic Oncology | Year: 2013
Objectives Epithelial ovarian cancer (EOC) is the major cause of death due to gynecological malignancies. The most important prognostic factors are residual tumor mass after surgery and platinum-response. No predictive biomarkers are available to identify patients who will benefit from standard treatment. The aim of our study was to analyze the role of HE4 in predicting surgical and clinical outcome in primary EOC. Methods In the European multicentric project "OVCAD", 275 consecutive patients with primary EOC were enrolled. Patients were eligible if radical cytoreductive surgery was performed and platinum-based chemotherapy was applied. Plasma and ascites samples were collected before or during surgery. The concentrations of HE4 and CA125 was determined using ELISA and Luminex technique, respectively. Results Median age at first diagnosis was 58 years (range 18-85 years). Most patients presented with advanced stage disease, FIGO III or IV (94.6%), grades II-III (96%) and serous histology (86.2%). In most cases a complete cytoreduction to no residual tumor mass was achieved (68.4%). Higher plasma HE4 levels correlated with poor surgery outcome in terms of macroscopically residual tumor mass (p < 0.001) and platinum-resistance (p = 0.009). Plasma CA125 and the risk index (HE4 and CA125) were independent predictive factors for surgical outcome (p = 0.001, OR = 3.37, 95% CI = 1.61-7.06 and p < 0.001, OR = 6,041, 95% CI = 2.33-15.65, respectively). FIGO stage III was an independent predictive factor for platinum response (p = 0.039, OR = 0.436, 95% CI = 0.198-0.960). Conclusions The presented data are showing that the combination of HE4 and CA125 expression in plasma might predict the surgical outcome in EOC and by this may have a prognostic impact on PFS and OS. © 2012 Elsevier Inc. All rights reserved.
Kaprio T.,University of Helsinki |
Fermer C.,Fujirebio Diagnostics AB |
Hagstrom J.,University of Helsinki |
Mustonen H.,University of Helsinki |
And 3 more authors.
BMC Cancer | Year: 2014
Background: Over two decades ago, a proposal was that two different colorectal cancer (CRC) entities existed, based on tumour location either proximal (right) or distal (left) of the splenic flexure. Proximal and distal tumours exhibit different clinical, epidemiological, and biological characteristics. Improvement of the prognostic evaluation of CRC requires new molecular markers. Podocalyxin-like 1 (PODXL), an anti-adhesive transmembrane sialomucin, is associated with an aggressive tumour phenotype and poor prognosis. For colorectal cancer, it has been suggested to be a marker of poor prognosis. The aim of this study was to investigate the role of PODXL in CRC by use of a novel monoclonal antibody.Methods: In 1983-2001, 840 consecutive colorectal cancer patients were treated at Helsinki University Central Hospital, of whom 767 were successfully scored for PODXL immunohistochemical expression from tumour tissue microarrays by use of a novel monoclonal in-house antibody. Associations of PODXL expression and tumour location with other clinicopathological variables were explored by Fisher's exact-test, linear-by- linear association test, and binary logistic regression. Survival analyses were done by the Kaplan-Meier method and Cox proportional hazards model.Results: PODXL protein expression was high in 44 (5.7%) specimens. High expression associated strongly with poor differentiation (p < 0.0001), advanced stage (p = 0.011), and location of the tumour in the right hemicolon (RHC) (p < 0.001). Tumours of the RHC were more poorly differentiated (p < 0.0001) and showed higher PODXL expression (p < 0.001).High PODXL expression associated significantly with higher risk for disease-specific death from CRC (hazard ratio (HR) = 2.00; 95% confidence interval (CI) 1.31-3.06, p = 0.001) and also in the subgroups of left hemicolon (LHC) cancers (HR = 2.60; 95% CI 1.45-4.66, p = 0.001) and rectal cancers (HR = 3.03; 95% CI 1.54-5.60, p = 0.001). Results remained significant in multivariable analysis (respectively, HR = 1.82; 95% CI 1.15-2.86, p = 0.01; HR = 2.59; 95% CI 1.41-4.88, p = 0.002; and HR = 2.69; 95% CI 1.30-5.54, p = 0.007).Conclusion: Podocalyxin was an independent factor for poor prognosis in colorectal cancer and in the subgroups of left hemicolon and rectum. This is, to our knowledge, the first evidence of such difference in PODXL expression, its function possibly being dependent upon tumour location. © 2014 Kaprio et al.; licensee BioMed Central Ltd.
Vucetic Z.,Fujirebio Inc. |
Dnistrian A.,Sloan Kettering Cancer Center |
Nilsson O.,Fujirebio Diagnostics AB |
Lilja H.G.,Sloan Kettering Cancer Center |
Plebani M.,University of Padua
Clinical Chemistry and Laboratory Medicine | Year: 2013
Background: Quality control materials with minimal inter-assay differences and clinically relevant proportions of different molecular forms of the analyte are needed to optimize intra- and inter-laboratory accuracy and precision. Methods: We assessed if clinically relevant total prostate- specific antigen (tPSA) levels were present in seven commercially available Multi Constituent Tumor Marker Controls (MC-TMC). Further, we determined the concentration of free PSA (fPSA) and calculated the percentage of free PSA (%fPSA) in all materials. Finally, we determined variability of TMC materials across several commonly used PSA platforms. Results: All MC-TMC materials contained at least one concentration of tPSA in normal and pathologic range. Control materials varied in the amount of fPSA and %fPSA, with most controls consisting of fPSA only and only one MC-TMC containing medically relevant levels of around 35% fPSA. Only a minority of MC-TMC materials showed minimal variability across four PSA methods while the majority of PSA controls showed wide inter-method differences. Conclusions: Use of many commercially available controls for PSA could lead to biased PSA measurements because they contain medically irrelevant proportions of fPSA and show significant variation among different PSA assay platforms.
Olofsson K.,Lund University |
Runquist D.,Lund University |
Runquist D.,Fujirebio Diagnostics AB |
Hahn-Hagerdal B.,Lund University |
Liden G.,Lund University
AMB Express | Year: 2011
Genetically engineered Saccharomyces cerevisiae strains are able to ferment xylose present in lignocellulosic biomass. However, better xylose fermenting strains are required to reach complete xylose uptake in simultaneous saccharification and co-fermentation (SSCF) of lignocellulosic hydrolyzates. In the current study, haploid Saccharomyces cerevisiae strains expressing a heterologous xylose pathway including either the native xylose reductase (XR) from P. stipitis, a mutated variant of XR (mXR) with altered co-factor preference, a glucose/xylose facilitator (Gxf1) from Candida intermedia or both mXR and Gxf1 were assessed in SSCF of acid-pretreated nondetoxified wheat straw. The xylose conversion in SSCF was doubled with the S. cerevisiae strain expressing mXR compared to the isogenic strain expressing the native XR, converting 76% and 38%, respectively. The xylitol yield was less than half using mXR in comparison with the native variant. As a result of this, the ethanol yield increased from 0.33 to 0.39 g g-1 when the native XR was replaced by mXR. In contrast, the expression of Gxf1 only slightly increased the xylose uptake, and did not increase the ethanol production. The results suggest that ethanolic xylose fermentation under SSCF conditions is controlled primarily by the XR activity and to a much lesser extent by xylose transport. © 2011 Olofsson et al; licensee Springer.
Fujirebio Diagnostics Ab | Date: 2015-02-04
The present invention provides a composition comprising at least two targeting agents, wherein at least one first targeting agent recognizes a keratin 7 peptide and/or fragment(s) thereof, and at least one second targeting agent recognizes a keratin 19 peptide and/or fragment(s) thereof. Said first and second targeting agents are capable of binding specifically and simultaneously to a heterotypic complex of keratin 7 with keratin 19, and/or fragment(s) thereof. The composition of the invention may be used in methods for diagnosing and/or prognosing, predicting efficacy of treatment, assessing outcome of treatment of assessing recurrence of malignant neoplastic disease, such as e.g. lung cancer, bladder cancer, esophagus cancer, and ovarian cancer in a subject.