Neste trabalho foram estudados diversos métodos cromatográficos para a purificação de fatores recombinantes de coagulação VII (FVIIr) e VIII (FVIIIr) derivados de linhagens celulares humanas SK-Hep. O FVIIIr é utilizado para o tratamento da Hemofilia A, enquanto o FVIIr é utilizado para o tratamento da Hemofilia B e também a Hemofilia A. Produzir estes fatores em linhagens celulares humanas faz com os padrões de glicosilação, sulfatação e enovelamento destas proteínas sejam extremamente parecidos com os fatores endógenos produzidos no organismo humano. A purificação do FVIIIr através de técnicas de cromatografia multimodais usando a resina CaptoMMC, afinidade usando a resina FVIIISelect e troca iônica (SP-Sepharose) permitiu obter um produto bastante homogêneo e com perfil de banda (por SDS-PAGE) bem definido que demonstrou a presença esperada das cadeias leve e pesada (o Westen-Blott indicou que os anticorpos comerciais reconheceram a cadeia pesada da molécula estudada). As técnicas permitiram uma alta reprodutibilidade do processo onde sequencias de purificação indicaram o mesmo comportamento de perfis cromatográficos e o processo eliminou 99.5% ± 0,5% de proteínas inespecíficas, recuperando até 64% de FVIIIr. O FVIIr foi purificado com apenas uma única técnica cromatográfica usando a resina FVIISelect que isolou a proteína de interesse eliminando cerca de 99% de impurezas, recuperando praticamente todo o produto. O eluido da cromatografia de afinidade foi dialisado em membranas de 5 kDa o que resultou no processo de auto ativação da molécula de FVIIr, resultando em um aumento de sinal de até 5x em relação a quantidade inicial. O gel de SDS-PAGE e o Westen-Blott comprovaram o processo de auto-ativação no qual uma migração de banda de 50 kDa para 30kDa foi observada e os anticorpos comerciais contra FVII foram capazes de detecta-la. O método de purificação também foi bastante reproduzível e o perfil de banda muito semelhante se comparado ao produto comercial existente no mercado. Sendo assim, foi possível obter plataformas de purificação para as proteínas FVIIr e FVIIIr. / In this work, several chromatographic methods were studied for the purification of recombinant clotting factors VII (FVIIr) and VIII (FVIIIr) derived from human SK-Hep cell lines. The FVIIIr is used for the treatment of Hemophilia A, while the FVIIr is used for the treatment of Hemophilia B and Hemophilia A. Producing these factors in human cell lines results in glycosylation, sulphation and folding patterns similar to the endogenous factors produced in the human organism. Purification of FVIIIr by multimodal chromatography techniques using CaptoMMC resin, affinity using FVIIISelect resin and ion exchange (SP-Sepharose) yielded a fairly homogeneous and well-defined band profile (by SDS-PAGE) which demonstrated the expected presence of the light and heavy chains, Westen-Blott indicated that commercial antibodies recognized the heavy chain of the studied molecule. The techniques allowed a high reproducibility of the process where purification sequences indicated the same behavior of chromatographic profiles and the process eliminated 99.5% ± 0.5% nonspecific proteins and recovering up to 64% FVIIIr. FVIIr was purified with only a single chromatographic technique using the FVIISelect resin which isolated the protein by removing about 99% impurities and recovering virtually the entire product. The affinity chromatography eluate was dialyzed on 5 kDa membranes which resulted in the autoactivation process of the FVIIr molecule resulting in a signal increase of up to 5 fold over the initial amount. The SDS-PAGE gel and Westen-Blott demonstrated the auto-activation process where a migration of 50 kDa to 30 kDa band was observed and the commercial antibodies against FVII were able to detect the band. The purification method was also quite reproducible and the band profile very similar compared to the commercial products. Thus, it was possible to obtain purification platforms for the FVIIr and FVIIIr proteins.

L’objet de notre réflexion est de voir que l’articulation entre le Conseil de sécurité et les juridictions pénales internationales s’appréhende essentiellement sous deux angles complètement nouveaux en droit international : la participation directe ou indirecte à la création d’organes juridictionnels et l’intrusion dans les procédures de fonctionnement de ceux-ci. Juridiquement, le Conseil de sécurité exerce pleinement ce double rôle sous le prisme de l’article 24 de la Charte des Nations Unies qui lui attribue la responsabilité principale en matière de maintien de la paix et de la sécurité internationales. En effet, qu’il s’agisse de la création des T.P.I., de son rôle peu ou prou variable dans l’établissement des juridictions mixtes, du pouvoir de saisine et de suspension de la C.P.I. que lui confère le Statut de Rome, de l’obligation de coopération des Etats dont il est l’ultime gardien ou de son pouvoir discrétionnaire dans la qualification du crime d’agression, le Conseil de sécurité est devenu, grâce à l’inépuisable fondement du Chapitre VII, le véritable catalyseur de la justice pénale internationale. Cependant, le revers de la médaille de cette importance du Conseil n’est pas anodin ou sans intérêt. Son irruption, très contestée au départ, dans le domaine de la justice pénale internationale donne lieu à une confrontation ancienne en droit international : celle de la politique et du juridique. Les termes de ce conflit transparaissent assez clairement de l’esprit de notre analyse. On ne peut d’ailleurs y échapper tant les préoccupations liées à la justice pénale internationale et celles relatives au maintien de la paix sont consubstantiellement imbriquées. Nous observons, non sans une certaine distance et une forme de retenue, que les termes mêmes de l’articulation alimentent toutes formes de conjectures sur l’indépendance et l’impartialité des juridictions pénales internationales. / The purpose of our research is to acknowledge that the link between the Security Council and international criminal jurisdictions is to be understood essentially under two entirely new perspectives in international law: the direct or indirect involvement of the Security Council in the creation of jurisdictional bodies and its intrusion in the operating procedures of the latter. Legally, the Security Council fully exercises this double role in accordance with Article 24 of the Charter of the United Nations, which devolves to it the main responsibility in the matter of maintaining peace and international security. Indeed, the Security Council, whether it is in the creation of ICCs, in its variable role in the setting up of mixed courts, in its power of submission and suspension of cases before the ICP which is imparted to it by the Statute of Rome, in its ultimate part in ensuring the obligation mutual cooperation amongst states, or in its discretionary power in the characterization of the crime of aggression, has become, thanks to the abiding foundation of Chapter VII, the genuine catalyst of international criminal justice. However, the downside to this importance of the Security Council is neither to be overlooked nor without interest. The sudden, and at first highly debated emergence of the Security Council in the area of international criminal justice has rekindled an old debate in international law: that of the political and the juridical order. The terms of this conflict show quite clearly through the approach of our analysis. We cannot evade this debate, considering how intricately the issues and concerns of international criminal justice and those of relating to peacekeeping are consubstantially interwoven. We may notice, albeit at some distance and some restraint, that the very terms of this link nurture all sorts of conjectures over the independence and impartiality of the international criminal courts.

A cours des dernières décennies, le Conseil de sécurité, agissant en vertu du chapitre VII de la Charte des Nations Unies, a autorisé l'adoption de diverses mesures de nature ou de portée économique, dans la sphère des États ou territoires non-étatiques, à l'issue de conflits armés. Il a ainsi promu la stabilisation économique, via des réformes économiques d'envergure, dans des zones post-conflictuelles telles que le Bosnie-Herzégovine, le Kosovo, le Timor oriental ou l'Irak. Il a intégré des principes de bonne gouvernance économique dans des régimes de gouvernance transitoires au Liberia ou aux deux Soudan. Il a développé une gamme de stratégies de consolidation de la paix de plus en plus sophistiquée pour stabiliser l'exploitation des ressources naturelles comme le bois, le pétrole ou les diamants, dans les États comme la Sierra Leone, la République démocratique du Congo ou la Côte d'Ivoire. Toutes ces interventions, plus ou moins intrusives, dans la sphère économique interne des États soulèvent des questions d'ordre juridique importantes. Dans cette étude nous nous sommes intéressés à deux questions essentiellement, à savoir, l'ordre juridique international, tel que nous le connaissons aujourd'hui, dispose-t-il de tous les matériaux nécessaires et indispensables pour encadrer et réglementer toute activité de reconstruction économique entreprise par le Conseil de sécurité ou ses agents d'exécution sur la base du chapitre VII d la Carte des Nations Unions ? Et si oui, comment et dans quelle mesure un tel cursus est-il appréhendé en pratique par les différents acteurs ? Les réponses apportées à cette problématique nous ont conduit, dans un premier temps, à déterminer l'existence , sinon la portée du corpus juridique pertinent, et ensuite, à analyser son impact sur la nature et la portée des pouvoirs susceptibles d'être exercé dans ce contexte. Et, dans un second temps, nous avons apprécié jusqu'à quel point les exigences ainsi établies ont, jusqu'à ce jour, pénétré la pratique des différents acteurs sur cette question précise, et en cas de violation des règles pertinentes, comment rendre, et dans quelle mesure, les acteurs concernés responsables des violations alléguées. / Résumé non communiqué

Šis darbas supažindina su tamsiojo turizmo fenomenu, kuris Lietuvos akademiniame pasaulyje dar nėra nagrinėtas. Darbe pristatoma tamsiojo turizmo samprata ir formų įvairovė, visuomenėje daug diskusijų keliantys šios turizmo krypties etiniai aspektai bei didėjančio populiarumo prielaidos. Taip pat iškeliama tamsiojo paveldo problematika, jį išskiriant kaip tam tikrą disonuojančio paveldo grupę. Darbe įvertinama dabartinė tamsiojo paveldo situacija Lietuvoje, išryškinant tvarkybos problemas ir lyginant tamsiojo paveldo pateikimo turizmui praktikas su teorinėje literatūroje rekomenduojamomis bei kritikuojamomis. Išsamiai analizuojamas gana ryškaus tamsiojo paveldo objekto – Kauno tvirtovės fortų – atvejis, įvertinant šio objekto reikšmę, dabartinę tvarkybą ir tai lemiančias priežastis. Kadangi iš visų devynių išlikusių Kauno tvirtovės fortų turizmui pritaikyti tik du (VII ir IX), kuriuose vystoma muziejinė veikla, įvertinamos tamsiojo paveldo reprezentacijos juose: tiriama šių dviejų Kauno tvirtovės fortų vykdoma veikla ir jos atitikimas užsienio autorių pateiktiems tamsiojo paveldo pritaikymo turizmui principams. / The paper introduces the new phenomena of dark tourism that has not ever been analyzed in Lithuanian academic literature. The notion of dark tourism and the variety of its forms is represented. This paper also analyzes the reasons of increasing popularity of dark tourism in postmodern society and the ethical points of dark tourism. This paper concentrates on assessment of Lithuania‘s dark heritage and explains the reasons, why this heritage is in such condition. The main focus is on Kaunas Fortress which is an outstanding example of dark heritage in Lithuania. The representation of dark heritage in Kaunas VII and Kaunas IX museums are compared and also the Lithuanian tourist‘s motivation of visiting the dark sites is analyzed.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

Le procédé PUREX utilise comme molécule extractante le tri-butyl-phosphate (TBP) pour assurer l’extraction et la purification de l’uranium et du plutonium des combustibles nucléaires usés. Bien que le retour d’expérience industriel soit très positif, des améliorations de ce procédé sont possibles notamment pour le futur traitement de combustibles plus riches en plutonium. Les extractants de la famille des N,N-dialkylamides (monoamides) ont montré par le passé une potentialité avérée pour remplacer le TBP : ils permettent de co-extraire l’uranium et le plutonium puis de dés-extraire sélectivement le plutonium par une diminution de l’acidité du milieu, supprimant ainsi les opérations de réduction du plutonium consommatrices en réactif potentiellement toxique (hydrazine). Le travail de recherche mené au cours de cette thèse vise à comprendre et modéliser l’extraction de certains produits de fission et actinides mineurs : césium, europium, américium, ruthénium et technétium, éléments qui ne doivent pas contaminer les matières recyclées. Les systèmes extractants choisis pour l’étude sont constitués des monoamides N,N-di (éthyl-2 hexyl) butanamide (DEHBA), N,N-di (éthyl-2 hexyl) isobutanamide (DEHiBA), du mélange de ces deux monoamides, de N-méthyl-N-octyl-(2-ethyl)hexanamide (MOEHA) et de TBP, extractant actuellement utilisé à l’usine AREVA de La Hague, dilués dans le tétrapropylène hydrogéné (TPH). Pour y parvenir, une démarche multi-échelle a été mise en place avec l’acquisition de données de distribution et de données thermodynamiques. Ces données ont servi de support pour élaborer des modèles permettant de simuler le comportement des différents éléments dans les cycles d’extraction actuel ou futur. En parallèle et lorsque les conditions chimiques des systèmes le permettaient, des études de spéciation des phases organiques ont été entreprises, afin d’obtenir des informations sur la stœchiométrie des complexes formés et sur les mécanismes mis en jeu au cours de l’extraction.De ces travaux peuvent se dégager un certain nombre de conclusions :- l’extraction des nitrates de césium, d’europium et d’américium par les monoamides est très faible et a pu être correctement modélisée,- l’extraction du technétium par les monoamides semble répondre au même mécanisme que dans le cas du TBP : le technétium est principalement co-extrait en phase organique en formant des complexes mixtes avec l’uranium. En effet, un anion pertechnétate se substitue à un anion nitrate dans les complexes formés entre l’extractant et le nitrate d’uranyle, selon un mode de coordination monodenté, conduisant à une extraction exacerbée du technétium. La modélisation de l’extraction du technétium par les monoamides a pu être améliorée en reconsidérant l’estimation des écarts à l’idéalité en phase aqueuse avec une nouvelle variation du coefficient d’activité de l’acide pertechnétique en solution binaire dans l’eau,- avec les monoamides comme avec le TBP, le ruthénium est faiblement extrait en phase organique mais cette fraction extraite, aussi faible soit elle, constitue une contamination résiduelle du solvant. Les données acquises, grâce à la mise au point d’un protocole de préparation de solutions simulées, sont représentatives du comportement du ruthénium dans les conditions d’extraction des procédés. Les modèles développés permettent de très bien reproduire le comportement du ruthénium dans les opérations d’extraction de procédés mis en œuvre à l’échelle pilote, sur des solutions de dissolution réelles,- une approche nouvelle pour le calcul des coefficients d’activité en phase organique a été développée. Elle explicite les écarts à l’idéalité liés à l’association en phase organique via les équilibres de complexation, mais aussi, et pour la première fois, les effets liés à la répulsion entre espèces en les considérant comme des sphères dures. / The PUREX process is a solvent extraction method dedicated to the reprocessing of irradiated nuclear fuel in order to selectively extract uranium(VI) and plutonium(IV) from fission products and minor actinides. The tri-n-butylphosphate (TBP) is used as the extractant in the organic phase. Within the frame of the development of Generation IV reactors, new liquid-liquid extraction processes are under development for the reprocessing of spent nuclear fuels. The N,N-dialkylamides (monoamides) already showed their potentiality as promising alternative extractant to TBP for nuclear fuel reprocessing: they are able to extract U(VI) and Pu(IV) selectively by adjusting the nitric acid concentration without using Pu(IV) reducing agents. This study aims at understanding and modeling the extraction of some fission products and minor actinides: cesium, europium, americium, ruthenium and technetium, which may occur as impurities in the organic phase. In the present study, the extraction properties of N,N-di (ethyl-2 hexyl) butanamide (DEHBA), N,N-di (ethyl-2 hexyl) isobutanamide (DEHiBA), as well as a mixture of these two monoamides, N-methyl-N-octyl-(2-ethyl)hexanamide (MOEHA) and TBP, the extractant currently used in the PUREX process at the La Hague plant, diluted in TPH, were studied. For that purpose, a multi-scale approach has been used to describe the extraction mechanisms combining two different descriptions. Distribution and thermodynamic data were first determined from batch experiments. Based on these data, thermodynamic models were developed and are able to predict the behaviour of the different elements in current or future processes. Dedicated methods were also performed to obtain information about the stoichiometry of the extracted species and about the mechanisms involved during the extraction step.The main conclusions that can be deduced from this study are:- even if the extraction of cesium, europium and americium nitrates with monoamides is very low, models have been developed and fit the experimental data with good agreement,- the same mechanism may be involved in the extraction of technetium with TBP or monoamides: technetium is preferentially co-extracted in organic phase as mixed uranium-technetium species. Indeed, one TcO4- anion replaces one NO3- ion in a monodentate coordination mode in the uranium-monoamide complex. The developed thermodynamic models, that have been improved by taking into account a new variation of the pertechnetic acid activity coefficient in binary solution, fit very well the experimental data,- with monoamides as with TBP, ruthenium is poorly extracted but remains troublesome in the spent fuel reprocessing industry because of its retention in the irradiated solvent. Distribution data have first been determined from batch experiments thanks to the development of a new methodology for simulated ruthenium spent fuel dissolution solutions preparation. The developed thermodynamic models fit very well the batch experimental data. Thus, they was then used to simulate ruthenium behaviour in counter-current hot tests performed in mixer-settlers- a new approach for the activity coefficient calculation in organic phase has been developed. The MSA theory (Mean Spherical Approximation) was chosen for this purpose to explicitly describe both association and repulsive forces.Finally, this work that includes a macroscopic study (distribution and thermodynamic data acquisition and modeling) and molecular investigations (ESI-MS, FT-IR and X-ray absorption analysis supported by theoretical calculations) provides a new insight in the description of solvent extraction mechanism.

EditorialThe text doesn't have an abstract / El texto no presenta resumen

Os experimentos com refletores nucleares de material pesado foram realizados no reator IPEN/MB-01 utilizando-se chapas de aço inoxidável, de aço carbono ou de níquel, num total de 32 chapas de cada material, inseridas adequadamente na face oeste do núcleo do reator. As chapas têm cerca de 3 mm de espessura. A largura e comprimento axial foram suficientes para cobrir todo o núcleo ativo do reator. Tais experimentos foram realizados com cada tipo de material refletor individualmente. Para cada etapa de colocação de chapas foram efetuadas medidas da reatividade devido à inserção destas no núcleo; bem como da posição crítica das barras de controle com BC1 e BC2 igualmente retiradas. Pôde ser observado que o aumento da absorção de nêutrons e consequente diminuição da moderação de nêutrons dominaram toda a física do problema quando foram inseridas poucas chapas de material refletor (cerca de 5 chapas para o aço inoxidável e aço carbono, e 3 chapas no caso do níquel). Na sequência, a reflexão de nêutrons tornou-se importante superando a absorção neutrônica; a reatividade aumentou até ultrapassar a situação sem chapa (excesso de reatividade zero) obtendo-se um acréscimo (ganho líquido) de reatividade com as 32 chapas inseridas (cerca de 162 pcm no caso do aço inoxidável, 37 pcm para o aço carbono e 295 pcm para o níquel). Portanto, observou-se que o núcleo refletido tornou-se mais reativo do que o núcleo sem material refletor. Resultados experimentais inéditos de medidas de reatividade foram obtidos com refletores de níquel. No que concerne a esse tipo de experimento não existe experimento similar na literatura internacional ao realizado no reator IPEN/MB-01. A análise teórica empregando o MCNP-5 e a biblioteca de dados nucleares ENDF/B-VII.0 evidenciou os aspectos físicos de absorção e reflexão de nêutrons nas chapas de material refletor considerados; entretanto apresentou uma discrepância quando a reflexão de nêutrons rápidos domina o fenômeno físico do transporte de nêutrons. Essas tendências foram encontradas independentemente do tipo de refletor pesado empregado nos experimentos. / Experiments with heavy-material nuclear reflectors were performed in the IPEN / MB-01 reactor using stainless steel, carbon steel or nickel plates, in a total of 32 plates of each material, properly inserted in the western face of the reactor core. The plates are about 3 mm thick. The axial width and length were sufficient to cover the entire active core of the reactor. Such experiments were performed with each type of reflective material individually. For each step of placement of plates were made measures of reactivity due to the insertion of these in the core; as well as the critical position of the BC1 and BC2 control rods also removed. It could be observed that the increase in neutron absorption and consequent decrease in neutron moderation dominated the whole physics of the problem when a few reflective material plates were inserted (about 5 plates for stainless steel and carbon steel and 3 plates for nickel). Subsequently, neutron reflection has become important overcoming neutron absorption; the reactivity increased until it exceeded the situation without plate (excess of zero reactivity) obtaining an increase (net gain) of reactivity with 32 inserts inserted (about 162 pcm in the case of stainless steel, 37 pcm for carbon steel and 295 pcm for nickel). Therefore, it was observed that the reflected core became more reactive than the core without reflective material. Unpublished experimental results of reactivity measurements were obtained with nickel reflectors. As for this type of experiment, there is no similar experiment in the international literature compared to the IPEN/MB-01 reactor. The theoretical analysis using the MCNP-5 together with nuclear data library ENDF/B-VII.0 evidenced the physical aspects of neutron absorption and reflection in the reflective material plates considered; however, it presented a discrepancy when the reflection of fast neutrons dominates the physical phenomenon of the transport of neutrons. These trends were found independently of the type of the heavy reflector employed in the experiments.

Le premier objectif de cette thèse est de réfléchir aux conditions de la genèse des grandes réformes universitaires à travers l’étude de la loi Faure de 1968 et de la loi Savary de 1984. Il s’agit d’essayer d’évaluer les rôles des différents acteurs, selon leur fonction et leur mode de pensée, dans le cadre du processus d’élaboration de la réforme. L’autre ambition assignée à ce travail est de tenter d’évaluer le degré d’application de la loi Faure de 1968 dans les universités. Une telle étude revêt une importance toute particulière dans la mesure où les historiens ont montré que cette loi est la première qui offre les moyens aux universités de devenir autonomes. Jusqu’à cette date, et malgré les réformes républicaines de la fin du XIXe siècle, l’enseignement supérieur français est resté tributaire de la « tyrannie des facultés » instituée par Napoléon en 1808. Nous avons donc cherché à évaluer le degré d’autonomie dont disposent les universités au lendemain de la loi de 1968 dans les domaines des diplômes, des finances et de la participation. Les sources autorisant la conduite d’une telle analyse sont très variées : dossiers des ministres en charge des universités, archives de l’Élysée, de différents rectorats, transcriptions des débats tenus à l’Assemblée nationale et au Sénat, rapport des commissions parlementaires, témoignages oraux a posteriori, etc... Pour ce qui concerne les sources émanant des universités, nous avons axé nos recherches sur les archives relatives à trois établissements : Paris VII, Rennes I et l’université de Picardie. / Our project strives for pondering the conditions for gradually implementing self-governance in the French universities. From this point of view however, the time frame spanning 1968 to 1984 is apparently a great opportunity for reflection. In fact the laws advocated by the Ministers Faure and Savary can be considered as the first steps towards practical autonomy of the universities.. History shows abundantly that despite republican reforms passed by the end of the 19th century, until 1968 the French higher education systems remained dependent on the “Colleges’ tyranny” established by Napoleon in 1808. A description of the French education system before 1968 will shed light on the administrative burdens on the universities, which may have aroused a willingness to change in some individuals. But in most cases, a reform derives from a trade-off whose origin may be tracked by historians. For such a quest, the sources of information flow from politicians of the time, or from their assistants, from academics or from teachers unions: proceedings of the Parliament or the Senate, reports from ministerial staffs, university bodies, ex post facto oral evidence etc…Confronting the material should allow one to gauge the part played in the reform preparation process by the various stakeholders, according to their function and mindset.

Neste trabalho foram estudados diversos métodos cromatográficos para a purificação de fatores recombinantes de coagulação VII (FVIIr) e VIII (FVIIIr) derivados de linhagens celulares humanas SK-Hep. O FVIIIr é utilizado para o tratamento da Hemofilia A, enquanto o FVIIr é utilizado para o tratamento da Hemofilia B e também a Hemofilia A. Produzir estes fatores em linhagens celulares humanas faz com os padrões de glicosilação, sulfatação e enovelamento destas proteínas sejam extremamente parecidos com os fatores endógenos produzidos no organismo humano. A purificação do FVIIIr através de técnicas de cromatografia multimodais usando a resina CaptoMMC, afinidade usando a resina FVIIISelect e troca iônica (SP-Sepharose) permitiu obter um produto bastante homogêneo e com perfil de banda (por SDS-PAGE) bem definido que demonstrou a presença esperada das cadeias leve e pesada (o Westen-Blott indicou que os anticorpos comerciais reconheceram a cadeia pesada da molécula estudada). As técnicas permitiram uma alta reprodutibilidade do processo onde sequencias de purificação indicaram o mesmo comportamento de perfis cromatográficos e o processo eliminou 99.5% ± 0,5% de proteínas inespecíficas, recuperando até 64% de FVIIIr. O FVIIr foi purificado com apenas uma única técnica cromatográfica usando a resina FVIISelect que isolou a proteína de interesse eliminando cerca de 99% de impurezas, recuperando praticamente todo o produto. O eluido da cromatografia de afinidade foi dialisado em membranas de 5 kDa o que resultou no processo de auto ativação da molécula de FVIIr, resultando em um aumento de sinal de até 5x em relação a quantidade inicial. O gel de SDS-PAGE e o Westen-Blott comprovaram o processo de auto-ativação no qual uma migração de banda de 50 kDa para 30kDa foi observada e os anticorpos comerciais contra FVII foram capazes de detecta-la. O método de purificação também foi bastante reproduzível e o perfil de banda muito semelhante se comparado ao produto comercial existente no mercado. Sendo assim, foi possível obter plataformas de purificação para as proteínas FVIIr e FVIIIr. / In this work, several chromatographic methods were studied for the purification of recombinant clotting factors VII (FVIIr) and VIII (FVIIIr) derived from human SK-Hep cell lines. The FVIIIr is used for the treatment of Hemophilia A, while the FVIIr is used for the treatment of Hemophilia B and Hemophilia A. Producing these factors in human cell lines results in glycosylation, sulphation and folding patterns similar to the endogenous factors produced in the human organism. Purification of FVIIIr by multimodal chromatography techniques using CaptoMMC resin, affinity using FVIIISelect resin and ion exchange (SP-Sepharose) yielded a fairly homogeneous and well-defined band profile (by SDS-PAGE) which demonstrated the expected presence of the light and heavy chains, Westen-Blott indicated that commercial antibodies recognized the heavy chain of the studied molecule. The techniques allowed a high reproducibility of the process where purification sequences indicated the same behavior of chromatographic profiles and the process eliminated 99.5% ± 0.5% nonspecific proteins and recovering up to 64% FVIIIr. FVIIr was purified with only a single chromatographic technique using the FVIISelect resin which isolated the protein by removing about 99% impurities and recovering virtually the entire product. The affinity chromatography eluate was dialyzed on 5 kDa membranes which resulted in the autoactivation process of the FVIIr molecule resulting in a signal increase of up to 5 fold over the initial amount. The SDS-PAGE gel and Westen-Blott demonstrated the auto-activation process where a migration of 50 kDa to 30 kDa band was observed and the commercial antibodies against FVII were able to detect the band. The purification method was also quite reproducible and the band profile very similar compared to the commercial products. Thus, it was possible to obtain purification platforms for the FVIIr and FVIIIr proteins.