An investigation into the folding and assembly of engineered antibodies and novel antibody formats

Monoclonal antibodies and novel antibody formats are currently one of the principal therapeutic in the biopharmaceutical industry worldwide and are widely used in the treatment of autoimmune diseases and cancer. It is for this reason that the productivity and quality of antibody production requires improvement; specifically investigations into the engineering of antibodies and any issues that may arise from the production of these therapeutics. The work presented in this thesis describes an investigation into the folding and assembly of seven antibodies plus the novel antibody format FabFv. IgG is comprised of two identical HCs and two identical LCs. The folding process of immunoglobulin is controlled by the CH1 domain within the HC. The CH1 domain remains in a disordered state and is sequestered by BiP in the endoplasmic reticulum. Upon the addition of a folded CL domain, BiP is displaced, the CH1 domain is able to fold and the complete IgG protein can then be secreted from the cell. The results presented in this thesis however, have outlined an additional mechanism for the folding of the CH1 domain. We have shown that the CH1 domain is able to fold in the absence of LC resulting in the secretion of HC dimers in a VH dependent manner. The proposed mechanism for the secretion of HC dimers suggests that some VH domains can interact with each other in order to bring the CH1 domains in close proximity to enable folding to occur. As HC dimer secretion is a hindrance in antibody production, this result has highlighted an engineering target to improve antibody yield. Examination of the folding of IgG4 with the variable region A33 has revealed the inability to secrete LC dimers, cleavage of the HC during expression and secretion of HC dimers in the Fab, FabFv and full length forms. The attributes described have also been shown to be variable region dependent. This has introduced a new concept that the variable domain is important in determining the expression and secretion of antibodies and their individual chains. Pulse chase and 2D gel electrophoresis analysis of the novel antibody format FabFv has revealed that the folding and expression of the LC and HC causes multimeric species of FabFv to be secreted, as opposed to the monomeric form which is the desired therapeutic. Our hypothesis is that this process occurs via a LC dependent mechanism. The proposed hypothesis suggests that further engineering to the LC could diminish the formation and secretion of FabFv multimers. The results from these investigations can be applied to increase the productivity of therapeutics and increase the biological understanding of the domain interactions of IgG during folding, assembly and secretion.

Perception of disease based on the level education and type of employment. Influence on adherence to treatment in young patients with hemophilia

SIN FINANCIACIÓN

EDUCAÇÃO E A ASSEMBLEIA DE DEUS NO BRASIL: COMPARAÇÕES, RESULTADOS, TENDENCIAS

O ensino teológico nas assembleias de Deus começou em meio à divisão de grupos, dos que defendiam o ensino formal e os do que não concordavam com a aplicação desta modalidade de ensino. No ano de 2011 a Assembleia de Deus fez 100 anos de existência, e na década de 2000 houve uma procura acentuada em cursos de teologia, por ser uma ferramenta necessária e obrigatória para o desenvolvimento do fiel dentro da instituição como ministro do evangelho, o que traduzindo seria o mesmo que a promoção dentro de uma empresa, subindo na hierarquia. A Assembleia de Deus despertou o interesse de pesquisadores, havendo muitos trabalhos desenvolvidos sobre a instituição. Mas falta acúmulo sobre como tem se dado a educação nas Assembleias de Deus. Um dos métodos de pesquisa foi o da pesquisa bibliográfica e as referências utilizadas na pesquisa são bem conhecidas no meio acadêmico como: Max Weber, Peter Berger, Paul Freston, Antônio Gouveia de Mendonça, entre outros. O levantamento desses trabalhos, relacionados à educação e a Assembleia de Deus no Brasil, possibilita comparações, oferecendo resultados e divisando tendências. Este trabalho apresenta, assim, uma breve história da Assembleia de Deus, fundação, implantação e desenvolvimento, propondo por onde realmente começou essa instituição eclesiástica e principalmente a classe social que teve maior participação em sua gênese na história brasileira. Após o levantamento histórico, destacamos a análise das pesquisas que correspondem às exigências desse trabalho. Procura-se, assim, encontrar subsídios para uma contribuição à Instituição, compreendendo os diferentes aspectos da educação na Assembleia de Deus, como também para uma melhor compreensão dessa instituição religiosa no contexto da sociedade brasileira.

Education and the historiography of Ibero-American independence: elusive presences, many absences

This contribution analyses recent historiographical tendencies in research in the field of education at the time of the political emancipation of the former Spanish and Portuguese colonies in Latin America. The article briefly presents the complex educational scene in Latin America on the eve of the movements for independence. Due to the revolutionary character of the process of independence, it identifies educational history as one of the most significant absences in the historiography of independence. Notwithstanding, education has certainly been addressed by historians of education, mostly focusing on the colonial or postcolonial period, while largely neglecting the two decades after 1808. This indicates both the divide prevalent between historians of education and historians of independence and the rather nationalistic conceptual frame of existing scholarship.

Assembly and disassembly of the Golgi Complex: two prodesses arranged in a cis to trans direction

We have studied the disassembly and assembly of two morphologically and functionally distinct parts of the Golgi complex, the cis/middle and trans cisterna/trans network compartments. For this purpose we have followed the redistribution of three cis/middle- (GMPc-1, GMPc-2, MG 160) and two trans- (GMPt-1 and GMPt-2) Golgi membrane proteins during and after treatment of normal rat kidney (NRK) cells with brefeldin A (BFA). BFA induced complete disassembly of the cis/middle- and trans-Golgi complex and translocation of GMPc and GMPt to the ER. Cells treated for short times (3 min) with BFA showed extensive disorganization of both cis/middle- and trans-Golgi complexes. However, complete disorganization of the trans part required much longer incubations with the drug. Upon removal of BFA the Golgi complex was reassembled by a process consisting of three steps: (a) exist of cis/middle proteins from the ER and their accumulation into vesicular structures scattered throughout the cytoplasm; (b) gradual relocation and accumulation of the trans proteins in the vesicles containing the cis/middle proteins; and (c) assembly of the cisternae, and reconstruction of the Golgi complex within an area located in the vicinity of the centrosome from which the ER was excluded. Reconstruction of the cis/middle-Golgi complex occurred under temperature conditions inhibitory of the reorganization of the trans-Golgi complex, and was dependent on microtubules. Reconstruction of the trans-Golgi complex, disrupted with nocodazole after selective fusion of the cis/middle-Golgi complex with the ER, occurred after the release of cis/middle-Golgi proteins from the ER and the assembly of the cis/middle cisternae.

Essays on economics of education and inequality

El documento resume los resultados de los tres ensayos sobre economía de la educación y de igualdad de oportunidades que se realizaron para el caso de Colombia.

The government of God desirable : a sermon delivered at Newark, N.J., October, 1808, during the session of the Synod of New-York and New-Jersey /

Mode of access: Internet.

Reflections on the works of God in nature and providence, for every day in the year /

Published in London in 1854 under title: Reflections on the being and attributes of God, and on his works, both in nature and providence.

The hand of God in history; or, Divine providence historically illustrated in the extension and establishment of Christianity.

"Some of the authors consulted": p. [vi]

God in his Providence: a comprehensive view of the principles and particulars of an active Divine Providence over man,--his fortunes, changes, trials, entire discipline as a spiritual being,

Mode of access: Internet.

The Clerk's Manual of Rules, Forms and Laws for the Regulation of Business in the Senate and Assembly

Title Varies: the Clerk's Manual of Rules, Forms and Laws for the Regulation of Business In the Senate and Assembly of the State of New York; Manual for the Use of the Legislature of the State of New York

Investigation of strike in steel industries : hearings before the Commmittee on Education and Labor, United States Senate, Sixty-sixth Congress, first session pursuant to S. Res. 202 on the resolution of the Senate to investigate the strike in steel industries.

The title page for Part 2 says "S. Res. 188, Instructing the Committee on Education and Labor of the United States Senate to investigatethe strike of the employees of the steel mills of the United States, and so forth and S. Res. 202, authorizing the Committee on Education and Labor, in its investigation of the steel strike, to hold hearings, to employ a stenographer, to require the attendance of witnesses and the production of papers, documents, and so forth, and prescribing penalties for the refusal of witnesses to attend or answer questions."

Editorial. Marketization and commodification of adult education

The editorial presents the articles of this issue on the topic of marketization and commodification of adult education of which some are conceptual and some empirical. (DIPF/Orig.)

Nuclear medicine technologist education and training in Europe: literature and web-based findings

Purpose - The education and training of a nuclear medicine technologist (NMT) is not homogeneous among European countries, which leads to different scope of practices and, therefore, different technical skills are assigned. The goal of this research was to characterize the education and training of NMT in Europe. Materials and methods - This study was based on a literature research to characterize the education and training of NMT and support the historical evolution of this profession. It was divided into two different phases: the first phase included analysis of scientific articles and the second phase included research of curricula that allow health professionals to work as NMT in Europe. Results - The majority of the countries [N=31 (89%)] offer the NMT curriculum integrated into the high education system and only in four (11%) countries the education is provided by professional schools. The duration in each education system is not equal, varying in professional schools (2-3 years) and high education level system (2-4 years), which means that different European Credit Transfer and Accumulation System, such as 240, 230, 222, 210 or 180 European Credit Transfer and Accumulation System, are attributed to the graduates. The professional title and scope of the practice of NMT are different in different countries in Europe. In most countries of Europe, nuclear medicine training is not specific and curriculum does not demonstrate the Nuclear Medicine competencies performed in clinical practice. Conclusion - The heterogeneity in education and training for NMT is an issue prevalent among European countries. For NMT professional development, there is a huge need to formalize and unify educational and training programmes in Europe.

Guidelines for the diagnosis and treatment of schistosomal myeloradiculopathy

Schistosomal myeloradiculopathy is the most severe and disabling ectopic form of Schistosoma mansoni infection. The prevalence of SMR in centres in Brazil and Africa that specialise in attending patients with non traumatic myelopathy is around 5%. The initial signs and symptoms of the disease include lumbar and/or lower limb pain, paraparesis, urinary and intestinal dysfunctions, and impotence in men. The cerebrospinal fluid of SMR patients shows an increase in protein concentration and in the number of mononuclear cells in 90% of cases; eosinophils have been reported in 40%. The use of magnetic resonance imaging is particularly valuable in the diagnosis of Schistosomal myeloradiculopathy. The exclusion of other myelopathies and systemic diseases remains mandatory. Early diagnosis and treatment with steroids and schistosomicides provide a cure for most patients, whilst delayed treatment can result in irreversible physical disabilities or death. To improve awareness concerning Schistosomal myeloradiculopathy amongst public health professionals, and to facilitate the control of the disease, the Brazilian Ministry of Health has launched a program of education and control of this ectopic form of schistosomiasis. The present paper reviews current methods for the diagnosis of SMR and outlines protocols for treatment of the disease.