El factor estimulador de colonias de granulocitos (G-CSF) es un medicamento que pertenece a un grupo de proteínas hematopoyéticas. Se obtiene por vía recombinante en el Centro de Ingeniería Genética y Biotecnología, de Cuba, desde el año 2000 y es comercializado como Hebervital. Se indica a pacientes con neoplasias afectados de neutropenia febril, con tratamiento mielosupresor, seguido de trasplante de la médula ósea, entre otros. Induce alteraciones en la actividad biológica de los neutrófilos maduros que pudieran aumentar la defensa del huésped en respuesta a patógenos, como bacterias y hongos. En este trabajo se realizó un análisis específico del G-CSF para determinar su pureza y caracterización; además, se demostró su posible comparabilidad con productosde otras firmas comerciales. El patrón de bandas de las muestras en la electroforesis, así como el porcentaje del área y los perfiles cromatográficos bajo la curva en la cromatografía líquida de alta resolución, fueron similares entre los diferentes productos. La digestión enzimática y la separación proteolítica de los péptidos generaron mapas peptídicos reproducibles y de elevada similitud con el Neupogen(AU)
Granulocyte Colony Stimulating Factor (G-CSF) is member of haemopoietic proteins group used in medication. Itis obtained by recombination technique since 2000 at the Center for Genetic Engineering and Biotechnology ofCuba, and it is commercialized as Hebervital. It is recommended for patients with malignant tumors, affected by febrile neutropenia, bone marrow suppressor treatment followed by bone marrow transplant, among others. It induces disorders in the biological activity of the mature neutrophils that could increase host defense in responseto pathogens like bacterial and fungal infections. The objective of this project was to perform a specific analysis todetermine G-CSF purity and characterization and to demonstrate its comparability with other commercial products. The sample bands pattern in SDS-PAGE, the area percentage and the chromatographic profiles under the curves,in reverse phase high performance liquid chromatographic were similar. The enzymatic digestion and separationby proteolysis of peptides generated reproduces peptide's maps of great similarity with Neupogen(AU)
Granulocyte Colony-Stimulating Factor/therapeutic use , Neutropenia/therapy , Fever/complications
Se caracterizó el lote de interferón gamma utilizado como material de referencia por diferentes técnicas analíticas y se demostró mediante los diferentes análisis de varianza realizados para cada una de las técnicas ensayadas, que el lote cumple con lo aceptado para ser usado como material de referencia. El estudio de homogeneidad realizado por la técnica de cromatografía líquida de alta eficacia en fase reversa demostró que el material de referencia posee el grado de homogeneidad requerido para su uso. El lote es estable por 2 años a 70 °C, según estudios previos realizados a los lotes de producción de materia prima activa de interferón gamma humano recombinante
Interferon-gamma , Reference Standards
Se caracterizó el lote de interferón gamma utilizado como material de referencia por diferentes técnicas analíticas y se demostró mediante los diferentes análisis de varianza realizados para cada una de las técnicas ensayadas, que el lote cumple con lo aceptado para ser usado como material de referencia. El estudio de homogeneidad realizado por la técnica de cromatografía líquida de alta eficacia en fase reversa demostró que el material de referencia posee el grado de homogeneidad requerido para su uso. El lote es estable por 2 años a 70 °C, según estudios previos realizados a los lotes de producción de materia prima activa de interferón gamma humano recombinante(AU)
Interferon-gamma/analysis , Reference Standards
Cytokines, such as interferons (IFN), underlie many immunological functions and are increasingly implicated in disease-related symptoms and pathology. In order to study the potential roles of IFN alpha and its antagonists in autoimmune phenomena, the sera from 89 patients (aged 15-95 years, 65 females) diagnosed as having myasthenia gravis (MG) (2 months to 34 years duration) were tested for the presence of natural anti-IFN alpha-2b auto-antibodies. Sera were screened for anti-IFN alpha-2b by a sandwich-type enzyme immunoassay system. Ten (11.2%) and 6 (6.7%) sera were identified that contained positive-competing and non-competing anti-IFN alpha-2b auto-antibodies, respectively. The MG sera were further analyzed by immunobloting against reduced IFN alpha-2b and for neutralizing anti-IFN alpha activity in an antiviral assay cells system. From tested EIA positive-competing sera, 5 were shown to be positive by immunoblot and 6 sera were found to contain neutralizing anti-IFN alpha-2b. Four of the 6 neutralizing anti-IFN alpha-2b sera came from patients with thymoma-associated MG. The sera were studied for linear epitope recognition on the IFN alpha-2b molecule by a solid phase binding assay, in which overlapping peptides homologous with the entire IFN alpha-2b sequence were separately synthesized on a nitrocellulose sheet. Peptides number 2 (residues 8-21), 3 (15-28), 6 (33-46), 10 (63-76), 15 (98-112), and 21 (141-154) were immunoreactive. Peptide 21 was apparently associated with antiviral activity, although peptide 21 has not been previously described as an immunogenic determinant on the IFN alpha-2b molecule. These results indicate that neutralizing anti-IFN alpha-2b is often present in MG, particularly in cases of thymoma-associated MG, and recognize a variety of epitopes on the IFN alpha-2b molecule, including those involved in its biological activity. Two groups of IFN epitopes were described associated with patient's age but not with diseases evolution.
Antibodies/immunology , Epitope Mapping , Epitopes/immunology , Interferon-alpha/immunology , Myasthenia Gravis/immunology , Adult , Age Factors , Aged , Aged, 80 and over , Female , Humans , Immunoblotting , Male , Middle Aged , Time Factors
