| Lung diseases are the lesions of the lung or a pulmonary manifestation of systemic diseases.With the aging of the population and the youthfulness of the disease,the number of patients with lung disease will increase significantly in the future.Lung cancer and lung infections pose a great threat to human life and health in particular.Current clinical diagnostic techniques for lung diseases are usually invasive or radiation-based,and fast and convenient non-invasive screening has not yet been achieved.In addition,chemotherapy,a common treatment for lung cancer,druginduced cardiotoxicity must be evaluated in advance.This dissertation is dedicated to the study of the design and manufacturing method of Love Wave sensor.A well-performing automated instrument based on this sensor was built and successfully applied to the non-invasive detection of potential lung disease biomarkers and the in vitro cardiotoxicity assessment of anti-lung cancer drugs.The detection technology based on the Love Wave sensor helps to achieve rapid noninvasive screening of lung disease and quantitative detection of cardiotoxicity of antilung cancer drugs.The main content and innovations of this dissertation are given as follows:1.A Love Wave sensor and its automated,portable detection instrument are developed to achieve highly sensitive and linear detection of mass and viscosity changes.In this study,a Love Wave sensor was designed and processed based on micro/nano-fabrication technology,and its highly sensitive linear response to the change of phase and insertion loss at the resonant frequency was experimentally demonstrated for mass deposition and surface viscosity,respectively.After that,a complete set of automated sampling and detection instrument was developed for the sensor and the related software was designed.The instrument provides the same function as the vector network analyzer in detecting the output signal of the Love Wave sensor,thus laying the foundation for the subsequent implementation of various biosensing applications.2.A Love Wave sensor for saliva detection was designed and prepared to achieve quantitative,highly sensitive and interference-resistant detection of Cyfra21-1,a lung cancer disease biomarker in saliva.In this study,a self-assembled monolayer was modified on the surface of the Love Wave sensor based on the cross-linking reaction of cysteamine and glutaraldehyde.Then,an immunosensor was constructed by the "sandwich" immunosandwich method,and a microfluidic chip was used to control the sample flow.Then a gold nanoparticlelabeled detection antibody was used to amplify the mass signal.Its performance on detecting Cyfra21-1 in complex samples was validated by spiking experiment in artificial saliva.The results showed that the linear detection range of Cyfra21-1 in artificial saliva was 15.625~125 ng/m L with the limit of detection of 15.312 ng/m L,demonstrating the utility of the proposed immunosensor in detecting salivary lung cancer markers,which is expected to be used for rapid and non-invasive lung cancer screening.3.A Love Wave immunosensor was for exhaled breath condensate detection was designed and prepared to achieve non-invasive and highly sensitive detection of Creactive protein,a biomarker of pulmonary infection,in exhaled breath.In this study,a device for simultaneous collection of exhaled volatile organic compounds and exhaled breath condensate samples was constructed and its stable and reliable sample collection performance was verified.After that,the surface modification method of the immunosensor was optimized and the effectiveness of the modification process was characterized by electrochemical cyclic voltammetry and impedence spectroscopy.Thus,a Love Wave immunosensor for CRP detection was successfully constructed.In addition,immunogold staining linearly amplified the mass signal of the sensor,and successfully achieved a highly sensitive detection of the pulmonary infection biomarker CRP.The results showed that the linear range for CRP was 5~50 ng/m L,and the limit of detection was 1.768 ng/m L.Finally,the spiking experiment in EBC samples demonstrated that the immunosensor can quantitatively detect CRP in complex salivary environment.4.A Love Wave biosensor based on cardiomyocytes was designed and prepared to achieve rapid detection and evaluation of cardiotoxicity of drugs for lung cancer treatment.In this study,an in vitro cardiac model was established on the surface of the Love Wave sensor using microfluidic chips and the cardiomyocyte cell line HL-1.The performance of this cell-based biosensor in detecting adhesion changes of cardiomyocyte population was subsequently validated using two drugs with opposite effects,verapamil and isoproterenol.Finally,the cardiotoxicity of an anti-lung cancer drug doxorubicin hydrochloride(DOX)on cardiomyocytes was monitored in real time using this cell-based biosensor,and the final state of the cells was verified in combination with optical microscopy and the CCK-8 cell viability kit.In addition,an effective normalized signal processing method was proposed to reduce the inherent variation of different sensors.The results showed that the normalized insertion loss was proportional to the logarithm of DOX concentration in the range of 0.1~10 μM and was consistent with the results of microscopy and the CCK-8 kit,demonstrating that the cell-based biosensor can be used for rapid quantitative detection of cardiotoxicity of anti-lung cancer drugs. |
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