Ovarian cancer is a high mortality disease where early stage detection may have significant survival benefits. Promising next-generation non-invasive, biomarker-based screening modalities involve longitudinal monitoring of serum biomarkers and multi-marker panel detection. Here, rapid, sensitive, precise and multiplexable diagnostic platforms can facilitate biomarker validation along with early detection and screening, and this work attempts to exploit the programmable bio-nano-chip (p-BNC) immunosensor to address these specific translational needs in ovarian cancer. First, the p-BNC was adapted for Cancer Antigen 125 (CA125) quantitation, the current FDA standard, with prominent implications in novel early detection and screening modalities. Antibody pairs binding to distinct epitopes on CA125 were identified and the p-BNC operating variables (incubation times, flow rates and reagent concentrations) were attuned to deliver optimal analytical performance (inter- and intra-assay precision of 1.2% and 1.9% and Limit-of-Detection (LOD) 1.0 U/mL), competitive with current gold standards, but with a short analysis time of 43 minutes. Further validation of the system with advanced stage patient sera (n=20) demonstrated good correlation with 'gold standard' ELISA (R² = 0.97). Next, the p-BNC was adapted for concomitant analysis of CA125 and Human Epididymis Protein 4 (HE4), a novel multiplexed biomarker panel for early detection and screening. The HE4 immunoassay was developed to perform optimally with the 'rate determining' CA125 assay. Cross-reactivity analysis demonstrated high specificity multiplexing. The dose-response curves for the multiplexed CA125 and HE4 immunoassays were congruous with their singleplex counterparts with respective LODs of 0.51 U/mL and 4.18 pM and a total analysis time of 44 minutes. A small pilot scale clinical study was conducted to discriminate between surgically confirmed patient sera (n=8) and corresponding age-matched healthy controls (n=8) utilizing the multiplexed p-BNC, interpreted with a risk of ovarian malignancy algorithm. Successful discrimination was achieved between the groups with Receiver Operating Characteristic (ROC) curve AUC (Area Under the Curve) values of 1.00, 0.984 and 1.00 respectively for CA125, HE4 and the composite marker combination. Taken together, the analytical and clinical performance, multiplexing capabilities and the short turn-around times on the p-BNC offer methodological advancements over current gold standard techniques, indicating strong promise for ovarian cancer diagnostics. / text

Ovarian cancer has very high mortality because it is hard to diagnosis in early stages. Many ovarian cancer biomarkers (such as HE4, CA 125) are available and had been suggested as potential tools for early cancer detection. However, early cancer detection using serological markers will only become widely used if a new generation of sensors that can be handled in a clinical setting can be developed. A detection technology that is promising for miniaturization and integration in biomedical sensing devices is based on the phenomenon of the extraordinary light transmission (EOT) through arrays of nanoholes on metal films. EOT is an increase in light transmission observed at certain wavelengths that satisfy the surface plasmon resonance (SPR) condition of the nanostructure. The position of this resonance is affected by surface adsorption phenomenon, which is the basis for the biosensor. In this dissertation, the detection of the HE4 biomarker was demonstrated using EOT. The EOT-based detection was compared to two state-of-the-art analytical methods (ELISA and commercial SPR). Based on our experiments, it was found that ELISA has lowest detection limit, around 0.5 ng/mL for that particular protein (HE4). The detection limits for the commercial SPR, around 0.13 μg/mL was comparable to the nanohole-based detection limit, around 1.76 μg/mL. In contrast to ELISA, the SPR-based methods were label free, more time efficient, and more environmental friendly. An extra advantage of the nanohole scheme was that multiple samples could be analyzed simultaneously and in real time. Adsorption kinetic experiments were also performed to evaluate the rate constants of the HE4 binding to a surface coated with anti-HE4. The adsorption equilibrium constant for the HE4 – anti-HE4 system was determined to be (4.3 ± 2.1) x 107 M-1. / Graduate / 0487

Epithelial ovarian cancer is the fifth leading cause of cancer-related deaths in the United States. However, the mortality rate is relatively high, due to in part to the cancer being in an advance stage at diagnosis, since it is often asymptomatic at the early-stage with a ~94% of five-year survival rate if it is diagnosed at a localized stage (stage 1). Early detection of cancer would likely improve the survival rate. Scientists are searching for novel promising methods to detect ovarian cancer at an asymptomatic early stage; also, the method is cheap and user-friendly despite there are various techniques for ovarian cancer detection. Cancer antigen 125 (CA125), a type of serum biomarker that elevates ~50% of women with early-stage and ~80% of women with advanced-stage, is used mostly for screening epithelial ovarian cancer. However, the lack of sensitivity and specificity are known to be the main drawback of CA125. Finding new potential biomarkers that diagnose cancer at a localized stage will significantly reduce the mortality rate. Human epididymis protein 4 (HE4) is such a biomarker that has a higher sensitivity and specificity compared to all other known biomarkers, and recently it has been approved by food and drug administration (FDA) for clinical applications.In this project, we developed sandwich-type micro particles immunoassay for sensitive detection of HE4 biomarker in plasma. Here, we cross-link elemental particles to a specific functional group of the targeted biomolecules based on a covalent and non-covalent linking chemistry to improve the sensitivity and the selectivity of biomarker detection in which Fe3O4 and SiO2 microparticles were used to conjugate and purify the antibody-antigen in a media. The purified assay with the microparticles was analyzed with laser-induced breakdown spectroscopy (LIBS) for detection and quantization analysis of the HE4 biomarker. Furthermore, along with LIBS, Raman, Fourier transform infrared (FT-IR), and UV- Vis spectroscopic techniques were utilized to understand the conjugation dynamic and confirm the conjugation process.

Structured abstract Study objectives: Ovarian carcinoma 1/ comparison of sensitivities among monitored markers CA 125, HE4, CA 19-9, CEA, TK, TPS, MonoTotal 2/ comparison of false positivity of markers CA 125 and HE4 3/ use of CA 125, HE4 and ROMA index in the diagnostics of ovarian carcinoma 4/ use of CA 125 and HE4 in the follow-up of ovarian cancer Endometrial carcinoma 1/ feasibility of use of biomarkers CA125 and HE4 in patients with endometrial cancer in pre- operative management Study design: Retrospective data analysis Settings: Department of Obstetrics and Gynecology, Medical Faculty and Teaching Hospital in Pilsen Patients and Methods: Ovarian cancer 1/ Sensitivity of markers CA 125, HE4, CA 19-9, CEA, TK, TPS, and MonoTotal was assessed in 266 patients - 19 with ovarian cancer and 247 with benign disorders. 2/ False positivity of markers CA125 and HE4 was evaluated in a total of 390 patients with benign diagnoses - 60 women with endometriosis, 70 pregnant patients, 67 patients with ascites, 60 with pleural effusion, 25 with cardiac failure , 80 with renal insufficiency and 28 with hepatic failure. 3/ As a part of this objective we evaluated 552 patients with abnormal pelvic abnormality - 30 women had a histologically confirmed malignant ovarian tumor. Other 522 women had a benign condition. The...

Structured abstract Study objectives: Ovarian carcinoma 1/ comparison of sensitivities among monitored markers CA 125, HE4, CA 19-9, CEA, TK, TPS, MonoTotal 2/ comparison of false positivity of markers CA 125 and HE4 3/ use of CA 125, HE4 and ROMA index in the diagnostics of ovarian carcinoma 4/ use of CA 125 and HE4 in the follow-up of ovarian cancer Endometrial carcinoma 1/ feasibility of use of biomarkers CA125 and HE4 in patients with endometrial cancer in pre- operative management Study design: Retrospective data analysis Settings: Department of Obstetrics and Gynecology, Medical Faculty and Teaching Hospital in Pilsen Patients and Methods: Ovarian cancer 1/ Sensitivity of markers CA 125, HE4, CA 19-9, CEA, TK, TPS, and MonoTotal was assessed in 266 patients - 19 with ovarian cancer and 247 with benign disorders. 2/ False positivity of markers CA125 and HE4 was evaluated in a total of 390 patients with benign diagnoses - 60 women with endometriosis, 70 pregnant patients, 67 patients with ascites, 60 with pleural effusion, 25 with cardiac failure , 80 with renal insufficiency and 28 with hepatic failure. 3/ As a part of this objective we evaluated 552 patients with abnormal pelvic abnormality - 30 women had a histologically confirmed malignant ovarian tumor. Other 522 women had a benign condition. The...

Thèse numérisée par la Direction des bibliothèques de l’Université de Montréal / Le travail présenté dans cette thèse se concentre sur la diffusion inélastique de neutrons de 14 MeV sur les premiers niveaux excités du carbone, de l'oxygène, et du lithium. Les mesures des sections efficaces différentielles ont été réalisées à des angles compris entre 30° et 130°. Le faisceau de deutérons, d'une énergie de 170 keV provenant de l'accélérateur Cockcroft-Walton, frappe une cible de tritium, produisant ainsi des neutrons de 14 MeV par la réaction T(d,n)He4. L'énergie des neutrons diffusés est déterminée par une méthode de temps de vol, utilisant la détection de la particule associée à la production du neutron pour connaître son temps de départ. Le temps de vol est converti en une impulsion de tension, qui est mesurée par un sélecteur d'amplitude à 100 canaux. La résolution temporelle du système est de 3 nanosecondes. Pour tous les niveaux excités observés, les courbes des sections efficaces différentielles inélastiques présentent une asymétrie par rapport à 90 degrés. Les résultats sont comparés aux prédictions du modèle d'interaction directe de Glendenning, et l'accord est satisfaisant pour tous les niveaux étudiés.

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