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Doctoral Thesis
DOI
10.11606/T.87.2016.tde-21102016-104835
Document
Author
Full name
Claudia Iwashita Verinaud
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Cheng, Elisabeth (President)
Azevedo, Anita Mitico Tanaka
Bresolin, Igor Tadeu Lazzarotto
Crespo, Joaquín Cabrera
Lebrun, Ivo
Title in Portuguese
Desenvolvimento de duas estratégias de purificação de IgM a partir de plasma humano.
Keywords in Portuguese
Cromatografia líquida
Hemoderivados
IgM
Imunoglobulinas
Abstract in Portuguese
Nesse trabalho foram estudadas 2 estratégias visando a purificação de IgM a partir de plasma humano. A primeira estratégia baseou-se em um processo desenhado para a fábrica de produção de hemoderivados do Instituto Butantan para a produção dos fatores de coagulação VIII e IX, albumina e IgG. Após 2 colunas cromatográficas, a IgM foi separada das 2 proteínas mais abundantes do plasma, a albumina e a IgG e o enriquecimento da IgM em relação a IgA e IgG foi bastante satisfatória. A segunda estratégia está inserida no processo de fracionamento de plasma desenvolvida em nosso laboratório. Através de um estudo sistemático em coluna de troca aniônica, obtivemos uma fração contendo IgG, uma contendo albumina, ambos praticamente puros e uma terceira contendo IgM e outras proteínas contaminantes. Os resultados obtidos indicam que a IgM pode ser purificada com sucesso empregando ambas as estratégias. Entretanto, para se obter um produto mais homogêneo, será necessário ainda adicionar uma ou mais etapas de purificação.
Title in English
Development of two strategies for the purification of human plasma IgM.
Keywords in English
Hemoderivatives
IgM
Immunoglobulins
Liquid cromatography
Abstract in English
In this work we studied two strategies for the purification human plasma IgM. The first strategy was based on a process designed for the hemoderivatives production factory of the Butantan Institute for the production of coagulation factors VIII and IX, albumin and IgG. After 2 chromatographies, IgM was separated from the 2 most abundant plasma proteins, that is, albumin and IgG and the achieved enrichment of IgM in relation to IgG and IGA was very satisfactory. The second strategy is part of a plasma fractionation process under development in our laboratory. Through a systematic study using an anionic exchange column we separated a fraction containing IgG, a second containing albumin, being both nearly pure and a third fraction containing IgM and other contaminating proteins. The obtained results indicate that IgM can be purified successfully by both strategies. However, to obtain a more homogeneous product, additional purification steps are necessary.
 
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Publishing Date
2016-10-24
 
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