Che-Chuan Yang

Time-Evolution contrast of target MRI using high-stability antibody functionalized magnetic nanoparticles: an animal model

In this work, high-quality antibody functionalized Fe3O4 magnetic nanoparticles are synthesized. Such physical characterizations as particle morphology, particle size, stability, and relaxivity of magnetic particles are investigated. The immunoreactivity of biofunctionalized magnetic nanoparticles is examined by utilizing immunomagnetic reduction. The results show that the mean diameter of antibody functionalized magnetic nanoparticles is around 50 nm, and the relaxivity of the magnetic particles is 145 (mMċs)-1. In addition to characterizing the magnetic nanoparticles, the feasibility of using the antibody functionalized magnetic nanoparticles for the contrast medium of target magnetic resonance imaging is investigated. These antibody functionalized magnetic nanoparticles are injected into mice bearing with tumor. The tumor magnetic-resonance image becomes darker after the injection and then recovers 50 hours after the injection. The tumor magnetic-resonance image becomes the darkest at around 20 hours after the injection. Thus, the observing time window for the specific labeling of tumors with antibody functionalized magnetic nanoparticles was found to be 20 hours after injecting biofunctionalized magnetic nanoparticles into mice. The biopsy of tumor is stained after the injection to prove that the long-term darkness of tumor magnetic-resonance image is due to the specific anchoring of antibody functionalized magnetic nanoparticles at tumor.

Clinic Applications in Assaying Ultra-Low-Concentration Bio-Markers Using HTS SQUID-Based AC Magnetosusceptometer

The HTS superconducting quantum interference devices (SQUID) magnetometers meets the requirement for the early-stage or hard-to-detect in-vitro diagnosis because of its ultrahigh sensitivity. For example, the demand for assaying the ultra-low-concentration biomarkers of tumors is always existing. It would be better to quantitatively detect the vascular endothelial growth factor (VEGF) at the concentration of 1 pg/ml for diagnosing the early-stage malignancy. In this work, by using the HTS SQUID-based ac magnetosusceptometer and biofunctionalized magnetic nanoparticles, the low-detection limit for VEGF is sub-pg/ml. Furthermore, the clear difference in the VEGF concentrations in sera was found between normal people and tumor patients. Another example to demonstrate the high sensitivity and high specificity of the immunoassay based on the HTS SQUID ac magnetosusceptometer is the detection of biomarkers for Alzheimers Disease. The biomarkers for Alzheimers Disease in plasma are very rare (around 1-100 pg/ml). It is hardly possible to assay the biomarkers in plasma. Here, it has been demonstrated that the HTS SQUID ac magnetosusceptometer can detect the biomarkers at very low concentrations ( ~ 1 pg/ml). Through the assays on the biomarkers in plasma of more than 100 people, the clinical accuracy is almost 90%. These results show the niches of clinical applications using the HTS SQUID ac magnetosusceptometer.

Molecule-assisted nanoparticle clustering effect in immunomagnetic reduction assay

Immunomagnetic reduction assay is used to quantitatively detect bio-molecules. Many reports show that the to-be-detected bio-molecular concentration dependent reduction in the alternative-current (ac) magnetic susceptibility of a reagent is governed by the logistic function, which is a four-parameter function. One of the parameters relates to the increase in the rate of the magnetic reduction signal when the concentration of to-be-detected bio-molecules is increased. Theoretically, this parameter is attributed to the clustering associations between to-be-detected bio-molecules and labeling particles in the reagent. In an immunomagnetic reduction assay, the bioactive labeling particles are anti-body-functionalized magnetic nanoparticles. However, there is no detailed information about the effect of the clustering associations on this parameter. In this work, the clustering association is manipulated by controlling the concentrations of anti-body-functionalized magnetic nanoparticles in the reagent. The exp...

Plasma Levels of Aβ42 and Tau Identified Probable Alzheimer’s Dementia: Findings in Two Cohorts

The utility of plasma amyloid beta (Aβ) and tau levels for the clinical diagnosis of Alzheimer’s disease (AD) dementia has been controversial. The main objective of this study was to compare Aβ42 and tau levels measured by the ultra-sensitive immunomagnetic reduction (IMR) assays in plasma samples collected at the Banner Sun Health Institute (BSHRI) (United States) with those from the National Taiwan University Hospital (NTUH) (Taiwan). Significant increase in tau levels were detected in AD subjects from both cohorts, while Aβ42 levels were increased only in the NTUH cohort. A regression model incorporating age showed that tau levels identified probable ADs with 81 and 96% accuracy in the BSHRI and NTUH cohorts, respectively, while computed products of Aβ42 and tau increased the accuracy to 84% in the BSHRI cohorts. Using 382.68 (pg/ml)2 as the cut-off value, the product achieved 92% accuracy in identifying AD in the combined cohorts. Overall findings support that plasma Aβ42 and tau assayed by IMR technology can be used to assist in the clinical diagnosis of AD.

Development for High-Accuracy In Vitro Assay of Vascular Endothelial Growth Factor Using Nanomagnetically Labeled Immunoassay

Nanomagnetically labeled immunoassays have been demonstrated to be promisingly applied in clinical diagnosis. In this work, by using antibody-functionalized magnetic nanoparticles and a high-temperature superconducting quantum interference device ac magnetosusceptometer, the assay properties for vascular endothelial growth factor (VEGF) in serum are investigated. By utilizing the assay method so-called immunomagnetic reduction, the properties of assaying VEGF are explored. In addition, the VEGF concentrations in serum samples of normal people and patients with either colorectal or hepatocellular cancer are detected. The experimental results show that the low-detection limit for assaying VEGF is 10 pg/mL, which is much lower than the clinical cut-off VEGF concentration of 50 pg/mL for diagnosing malignancy. Besides, there are no significant interference effects on assaying VEGF from hemoglobin, conjugated bilirubin, and triglyceride. The VEGF concentrations in serum samples donated by normal people and patients with hepatocellular carcinoma or colorectal cancer are detected. A clear difference in VEGF concentrations between these two groups is found. These results reveal the feasibility of applying nanomagnetically labeled immunoassay to clinics.

Analytical performance of reagent for assaying tau protein in human plasma and feasibility study screening neurodegenerative diseases

Immunomagnetic reduction (IMR), which involves the use of antibody-functionalized magnetic nanoparticles to specifically label target biomarkers, was utilized to develop an assay for total tau protein in human plasma. The analytic properties of the IMR assay on tau protein were investigated. The limit of detection was found to be 0.026 pg/ml. Other properties such as Hook effect, assay linearity, dilution recovery range, reagent stability, interference test, and spiked recovery were also characterized. The ultra-sensitive IMR assay was applied to detect the plasma tau protein levels of subjects with prevalent neurodegenerative diseases, such as Alzheimer’s disease (AD), mild cognitive impairment (MCI) due to AD, Parkinson’s disease (PD), frontotemporal dementia (FTD) and vascular dementia (VD). The concentrations of plasma tau protein in patients with VD, PD, MCI due to AD, FTD, and AD patients were higher than that of healthy controls. Using an ROC curve analysis, the cutoff value for discriminating dementia patients from healthy controls was 17.43 pg/ml, resulting in 0.856 and 0.727 for clinical sensitivity and specificity, respectively. The area under the ROC curve was 0.908. These results imply that the IMR plasma tau assay would be useful to screen for prevalent neurodegenerative diseases.

Step-Edge High-

The fabrication processes for step-edge high- Tc superconducting-quantum-interference-device (SQUID) magnetometers have been developed. A magnetometer consists of three SQUIDs, which can be activated either individually or in series. Thus, the transfer function can be manipulated by activating a single SQUID or multiple SQUIDs in series. Furthermore, such a SQUID magnetometer is benefited by that if one of the three SQUIDs is broken, the other two SQUIDs are still workable. In addition to characterizing the SQUID magnetometer, the application of the SQUID magnetometer in the low-field nuclear magnetic resonance (NMR) has also been investigated. The results has shown a clear NMR signal of 4230 Hz for the water under 99.3 μT.

{\rm T}_{\rm c}

The 42 amino acid form of amyloid-β (Aβ42) plays a key role in the pathogenesis of Alzheimers disease (AD) and is a core biomarker for the diagnosis of AD. Numerous studies have shown that cerebrospinal fluid (CSF) Aβ42 concentrations are decreased in AD, when measured by enzyme-linked immunosorbent assay (ELISA) and other conventional immunoassays. While most studies report no change in plasma Aβ42, independent studies using the immunomagnetic reduction (IMR) technique report an increase in plasma Aβ42 levels in AD. To confirm the opposite changes of Aβ42 levels in CSF and plasma for AD, we assayed the levels of Aβ42 in plasma of subjects with known CSF Aβ42 levels. In total 43 controls and 63 AD patients were selected at two sites: the VU University Medical Center (n = 55) and Sahlgrenska University Hospital (n = 51). IMR and ELISA were applied to assay Aβ42 in plasma and CSF, respectively. We found a moderately negative correlation between plasma and CSF Aβ42 levels in AD patients (r = -0.352), and a weakly positive correlation in controls (r = 0.186). These findings further corroborate that there are opposite changes of Aβ42 levels in CSF and plasma in AD. The possible causes for the negative correlation are discussed by taken assay technologies, Aβ42 transport from brain to peripheral blood, and sample matrix into account.

SQUID Magnetometer for Low-Field NMR Detection

Objective: Parkinson’s disease (PD) has significant clinical overlaps with atypical parkinsonism syndromes (APS), which have a poorer treatment response and a more aggressive course than PD. We aimed to identify plasma biomarkers to differentiate PD from APS. Methods: Plasma samples (n = 204) were obtained from healthy controls and from patients with PD, dementia with Lewy bodies (DLB), multiple system atrophy, progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), or frontotemporal dementia (FTD) with parkinsonism (FTD-P) or without parkinsonism. We measured plasma levels of α-synuclein, total tau, p-Tau181, and amyloid beta 42 (Aβ42) by immunomagnetic reduction-based immunoassay. Results: Plasma α-synuclein level was significantly increased in patients with PD and APS when compared with controls and FTD without parkinsonism (p < 0.01). Total tau and p-Tau181 were significantly increased in all disease groups compared to controls, especially in patients with FTD (p < 0.01). A multivariate and receiver operating characteristic curve analysis revealed that a cut-off value for Aβ42 multiplied by p-Tau181 for discriminating patients with FTD from patients with PD and APS was 92.66 (pg/ml)2, with an area under the curve (AUC) of 0.932. An α-synuclein cut-off of 0.1977 pg/ml could separate FTD-P from FTD without parkinsonism (AUC 0.947). In patients with predominant parkinsonism, an α-synuclein cut-off of 1.388 pg/ml differentiated patients with PD from those with APS (AUC 0.87). Conclusion: Our results suggest that integrated plasma biomarkers improve the differential diagnosis of PD from APS (PSP, CBD, DLB, and FTD-P).

Plasma Amyloid-β (Aβ 42) Correlates with Cerebrospinal Fluid Aβ 42 in Alzheimer’s Disease

from ageand sex-matched controls, and from individuals with other neurodegenerative diseases. Results: Results demonstrate that detection of depleted autoantibodies in serum can readily differentiate disease subjects from ageand sex-matched controls with high overall accuracy, sensitivity, and specificity. Depletion biomarkers were able to distinguish subjects with early AD pathology at MCI from controls with 90.0% overall accuracy in the Testing Set (sensitivity1⁄488.0%; specificity1⁄492.0%). Comparable accuracies were obtained for identification of subjects with mild-moderate AD, early-stage Parkinson’s disease and multiple sclerosis. Conclusions: Results demonstrate that monitoring the disease-linked depletion of brain-reactive autoantibodies in blood has utility for early detection and staging of AD and other neurodegenerative diseases. Since autoantibody depletion may reflect neuropathological events prior to significant neurodegeneration, we hypothesize that depleted autoantibody biomarkers may be particularly useful for preclinical disease detection, perhaps years before the onset of symptoms.