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Master's Dissertation
DOI
10.11606/D.42.2014.tde-20022015-165502
Document
Author
Full name
Ângela Alice Amadeu Megale
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Silva, Wilmar Dias da (President)
Carvalho, Enéas de
Isaac, Lourdes
Title in Portuguese
Desenvolvimento de anticorpos anti-peptídeos sintéticos derivados da proteína transportadora de fosfato dependente de sódio NaPi2b.
Keywords in Portuguese
Anticorpos
Câncer ovariano
Moléculas carreadoras
NaPi2b
Peptídeos
Proteína
Abstract in Portuguese
Dois peptídeos, designados Let#1 e Let#2, foram delineados do segundo laço extracelular da proteína NaPi2b. Após conjugação, os peptídeos induziram produção de anticorpos específicos em coelhos e camundongos. Os anticorpos com maiores títulos foram selecionados para ensaios complementares. Estes anticorpos reconheceram os peptídeos conjugados aos carreadores pelos métodos de ELISA e WB, não havendo reação cruzada quando testados com a proteína carreadora livre e com antígeno sintético inespecífico. Após a completa validação, foi possível observar que os anticorpos anti-peptídeos reconhecem NaPi2b nativa, presente na superfície de células OV-CAR, bem como mostraram reconhecimento de um alvo comum em todos os soros analisados, mesmo quando o soro era considerado negativo para COV pelo teste comercial. Este reconhecimento não foi observado quando as amostras foram analisadas pelo soro pré-imune dos animais teste, sugerindo um reconhecimento específico dos anticorpos produzidos.
Title in English
Development of anti-synthetic peptides antibodies derived from sodium-dependent phosphate transporter NaPi2b protein.
Keywords in English
Antibodies
Carriers molecules
NaPi2b
Ovarian cancer
Peptides
Protein
Abstract in English
Two peptides, designated Let#1 and Let#2, were outlined from the second extracellular loop of the NaPi2b protein. After conjugation, the peptides induced the production of specific antibodies in rabbits and mice. Antibodies with higher titers were selected for complementary tests. These antibodies recognized peptides conjugated to the carrier by ELISA and WB, with no cross-reaction when tested with the free carrier protein and nonspecific synthetic antigen. After complete validation, it was observed that the anti-peptide antibodies recognize native NaPi2b protein present on the OV-CAR cell surface and showed recognizing a common target in all sera tested, even when the serum was considered negative for ovarian carcinonoma by commercial test. This recognition was not observed when the samples were analyzed by pre-immune serum of test animals, suggesting specific recognition of antibodies produced.
 
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Publishing Date
2015-02-24
 
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