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Doctoral Thesis
DOI
https://doi.org/10.11606/T.99.2019.tde-11022019-090657
Document
Author
Full name
Pâmela Soto Garcia
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Leite, Fabio de Lima (President)
Domingues, Renan Barros
Goto, Hiro
Silva, Marcelo de Assumpcao Pereira da
Title in Portuguese
Desenvolvimento de nanoimunossensores de microscopia de força atômica para estudo da esclerose múltipla
Keywords in Portuguese
Bainha de mielina
Esclerose múltipla
Espectroscopia atômica
Microscopia de força atômica
Sensor
Sistema nervoso central
Abstract in Portuguese
A glicoproteína oligodendrocítica da mielina (MOG) e proteína básica da mielina (MBP) têm sido implicadas como os antígenos-alvo mais importantes nos processos desmielinizantes do sistema nervoso central (SNC), e mais importantes autoantígenos que surgiram dos estudos com o modelo animal para a esclerose múltipla (EM), a encefalomielite autoimune experimental (EAE). Os primeiros autoanticorpos detectados no soro e liquido cefalorraquidiano (LCR) de pacientes com EM foram anticorpos contra antígenos da mielina. O diagnóstico diferencial da EM inclui a presença de bandas oligoclonais (BOCs) no LCR e ausência no soro, demonstrando dessa forma síntese intratecal de imunoclobulinas G (IgG). As técnicas de detecção de anticorpos mais utilizadas atualmente são ELISA, ensaio baseado em células e western blot (WB). Neste contexto, o estudo da anti-MOG e anti-MBP e seu papel na EM podem ser estudados através da técnica de espectroscopia de força atômica (AFS). Esta é uma técnica altamente sensível que permite a detecção molecular, com a interação de uma ponta funcionalizada de microscópio de força atômica (AFM) com uma amostra, a qual fornece desta forma a força de adesão (Fad) específica para o sistema. Nesta pesquisa, foi inserido na ponta de AFM funcionalizada os peptídeos encefalitogênicos MOG92-106 e MBP85-99, para detectar e estudar os anticorpos específicos IgG anti- MOG92-106 e MBP85-99, no soro e LCR de pacientes na amostra, utilizando a técnica AFS. Sendo assim, este estudo foi realizado de forma inédita utilizando a AFS, auxiliando diretamente na investigação da EM e doenças desmielinizantes relacionadas.
Title in English
Development of atomic force microscopy nanoimmunosensor applied to the survey of multiple sclerosis
Keywords in English
Atomic force microscopy
Central nervous system
Force spectroscopy
Multiple sclerosis
Myelin sheath
Sensor
Abstract in English
Myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) have been implied as the most important target antigens in demyelinating processes of central nervous system (CNS) and the most important antigens candidates whom arised from the animal model for multiple sclerosis (MS), the experimental autoimmune encephalomyelitis (EAE). The first autoantibodies detected in serum and cerebrospinal fluid (CSF) of MS patients were antibodies against myelin antigens. Differential diagnostic to MS includes the presence of oligoclonal bands (OCBs) in CSF and absence in serum, which demonstrate intrathecal IgG synthesis. Most applied techniques to detection of antibodies nowadays are ELISA, cell-based assay and western blot (WB). In this context, the study of anti-MOG role in e disease may be supported through its detection by atomic force spectroscopy (AFS) technique. AFS is a highly sensitive technique that allows molecular detection as a functionalized atomic force microscope (AFM) tip interacts with the sample, providing the system specific adhesion force (Fad). In this research, it was attached in the functionalized AFM tip the notable encephalitogenic peptides MOG92-106 and MBP85-99 to detect and study the specific antibodies anti-MOG92-106 and anti-MBP85-99 on the sample with AFS technique. Thus, this study was applied for the first time in research with AFS, assisting directly to MS and other demyelinating diseases investigation.
 
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Publishing Date
2019-02-11
 
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