Monoclonal antibody

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EL TRATAMIENTO CON EL ANTICUERPO MONOCLONAL CHP3R99 DISMINUYE LA INFILTRACIÓN DE MACRÓFAGOS EN LA ÍNTIMA ARTERIAL DE CONEJOS NZB / THE TREATMENT WITH THE MONOCLONAL ANTIBODY CHP3R99 REDUCES THE MACROPHAGE INFILTRATION WITHIN ARTERIAL INTIMA OF NZW RABBITS

EL TRATAMIENTO CON EL ANTICUERPO MONOCLONAL CHP3R99 DISMINUYE LA INFILTRACIÓN DE MACRÓFAGOS EN LA ÍNTIMA ARTERIAL DE CONEJOS NZB / THE TREATMENT WITH THE MONOCLONAL ANTIBODY CHP3R99 REDUCES THE MACROPHAGE INFILTRATION WITHIN ARTERIAL INTIMA OF NZW RABBITS

The retention of low-density lipoproteins (LDL) by arterial proteoglycans represent a key molecular event in atherosclerosis development. This process promotes the oxidation of LDL and the development of a chronic inflammatory response, leading to atherosclerosis progression. At the Center of Molecular Immunology, it was obtained a chimeric monoclonal antibody, named chP3R99, which recognizes sulfated proteoglycans of the arterial wall, and able to inhibit the retention and oxidation of LDL. In addition, the immunization with this antibody prevents atherosclerotic lesions development by mean a generation of anti-proteoglycans autologous antibodies. Nonetheless, the molecular mechanisms of action of this antibody remain in part unknown. Thus, the present work was aimed to evaluate the effect of chP3R99 immunization on the macrophage infiltration, because of the key role of this cellular population during vascular inflammation. We carried out a preventive immunization where we administered a dose of 100 µg, s.c. in rabbits later treated with 2 mL/kg i.v. of Lipofundin, an inductor of atherosclerotic lesions. The results showed that the treatment with the chP3R99 antibody inhibited the macrophage infiltration, at the same time that reduced the gene expression of inducible nitric oxide synthase and myeloperoxidase activity. These results demonstrate the potential of chP3R99 immunization to reduce the inflammatory response, a key factor in atherosclerosis development.

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DISPOSICIÓN FARMACOCINÉTICA DEL ANTICUERPO MONOCLONAL QUIMÉRICO P3 EN CONEJOS: CINÉTICA NO LINEAL / PHARMACOKINETIC DISPOSITION OF THE CHIMERIC MONOCLONAL ANTIBODY P3 IN RABBITS: NON LINEAR KINETICS

DISPOSICIÓN FARMACOCINÉTICA DEL ANTICUERPO MONOCLONAL QUIMÉRICO P3 EN CONEJOS: CINÉTICA NO LINEAL / PHARMACOKINETIC DISPOSITION OF THE CHIMERIC MONOCLONAL ANTIBODY P3 IN RABBITS: NON LINEAR KINETICS

El anticuerpo monoclonal quimérico (AcM quimérico) P3 fue obtenido en los laboratorios del Centro de Inmunología Molecular (CIM) de Ciudad de La Habana a través de técnicas de ingeniería de anticuerpos a partir del AcM P3 murino y mantuvo sus principales propiedades de reconocimiento, reaccionando específicamente con NeuGcGM3 y NeuGcGM2, pero no con sus variantes acetiladas. El producto ensayado fue formulado como solución inyectable a una concentración de 10 mg/mL en solución salina tamponada (PBS), teniendo en cuenta las Buenas Prácticas de Producción establecidas en los Procedimientos Normalizados de trabajo de dicha institución.

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Preclinical Diagnosis of Ovine Scrapie by Immunohistochemistry of Lymphoid Tissue Using a Panspecific Monoclonal Antibody Cocktail

Preclinical Diagnosis of Ovine Scrapie by Immunohistochemistry of Lymphoid Tissue Using a Panspecific Monoclonal Antibody Cocktail

Scrapie is a naturally occurring prion-associated disease of sheep and goats. A major impediment to research and control of scrapie has been the lack of a method to predict infection prior to the onset of clinical disease in live sheep or in slaughter surveillance. The scrapie-associated prion isoform PrP-Sc is detectable in lymphoid tissues, including those of the third eyelid (nictitating membrane) and tonsil, from infected sheep months or years before development of clinical disease. Third eyelid tissue can be collected using topical anesthesia and is suitable for screening live animals over 18 months of age for scrapie. In this study, we show a high concordance between the third eyelid preclinical test for ovine scrapie and the current diagnostic standards of spongiform lesions and/or immunohistochemistry of the medulla at the level of the obex. The results of third eyelid assay agreed with the scrapie status of sheep in 251 of 258 sheep sampled, including 27 sheep that progressed to clinical disease with confirmed scrapie 3 to 20 months following biopsy. Tonsil was positive for PrP-Sc at post mortem analysis in all sheep with immunostaining of brain. Standardized protocols for high throughput automated immunostaining and for manual staining with commercially available reagents were developed. Lymphoid based testing for PrP-Sc was performed with MAb F89/160.1.5, which binds residues 142-145 of ovine PrP, and MAb F99/97.6.1, which binds residues 220-225. One or both monoclonal antibodies in this cocktail recognize PrP sequences conserved in most mammalian species in which natural TSEs have been reported. The third eyelid test on live sheep and addition of tonsil tissue to postmortem or slaughter surveys will be useful in diagnostic, surveillance and research programs.

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CINÉTICA EN ESTADO DE ENFERMEDAD DEL ANTICUERPO MONOCLONAL QUIMÉRICO P3: MODELO ANIMAL DE ATEROSCLEROSIS / KINETICS ON STATE DISEASE OF THE CHIMERIC MONOCLONAL ANTIBODY P3: ANIMAL MODEL OF ATHEROSCLEROSIS

CINÉTICA EN ESTADO DE ENFERMEDAD DEL ANTICUERPO MONOCLONAL QUIMÉRICO P3: MODELO ANIMAL DE ATEROSCLEROSIS / KINETICS ON STATE DISEASE OF THE CHIMERIC MONOCLONAL ANTIBODY P3: ANIMAL MODEL OF ATHEROSCLEROSIS

El anticuerpo monoclonal murino P3, demostró su reconocimiento por los glicolípidos sulfatados y los proteoglicanos 12 , aunque su variante quimérica no la manifiesta con igual intensidad pero su identificación si ha posibilitado a través de la Ingeniería Genética, el diseño de novedosas estructuras derivadas para alcanzar las propiedades requeridas que inciden en una estrategia clínica orientadas hacia las alternativas terapéuticas a emplear en el tratamiento de la aterosclerosis. Cada año se incrementa el número de investigaciones que determinan el carácter autoinmune de las enfermedades, entre ellas la propia aterosclerosis. Ello ha definido novedosos conceptos terapéuticos que utilizan los mecanismos idiotípicos de diversos anticuerpos, para generar anticuerpos naturales que contrarresten la enfermedad 13, 14 .

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P3, un anticuerpo monoclonal capaz de activar clulas B 1a

P3, un anticuerpo monoclonal capaz de activar clulas B 1a

P3 is a monoclonal antibody (mAb) of IgM isotype, which recognizes N-glycolylated gangliosides and sulfatides, both self-antigens in mice. It was also re- ported that the variable region of P3 is shared by the antibody A4ac, isolated from a mouse with experi- mental allergic encephalomyelitis, which recognizes myelin oligodendrocytes [1]. The P3 mAb triggers a strong anti-idiotypic response in the syngeneic BALB/c mice model, even in the absence of adjuvant or carrier protein [2], which is not a common phenom- enon [3]. Some authors have suggested that the IgM isotype or the presence of somatic hypermutations could be important factors to explain the immuno- genicity of autologous immunoglobulins [4-6]. Cu- riously, the immunogenicity of the P3 mAb idiotype has been demonstrated in the absence of any constant domain [7] and the P3 mAb variable region is coded by germline genes [8]. Therefore, the intrinsic proper- ties of the P3 mAb idiotype and its capacity to interact with immune cells could explain the capacity of P3 to induce a strong anti-idiotypic response.

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Nimotuzumab, inmunoterapia eficaz para el tratamiento de tumores epiteliales malignos

Nimotuzumab, inmunoterapia eficaz para el tratamiento de tumores epiteliales malignos

After registering the nimotuzumab, an observational, prospective, multicenter and open clinical trial was carried out with the treatment of 577 patients with ad- vanced tumors of epithelial origin; 89 of them were of pediatric ages and 488 adults. In 19.1 and 22.5 %, of them respectively, there was at least one adverse event observed during the treatment. The association be- tween the exposure time to the monoclonal antibody, the number of doses applied and the frequency of the presence of adverse events was not proven. There was no influence in the intensity of the events either, regard- less the causal relationship. Most of the events were classified as mild or moderate. Very few severe adverse events were notified, i.e., one patient with anaphy- laxis, one with venous thrombosis, one with lipothy- mia and another one with tumoral lysis syndrome [37]. The overall report on nimotuzumab safety (issued in 2012) shows that out of a total of 38 629 patients, 10 % of them were treated in clinical trials and the other 90 % were treated following the doctor’s pre- scription. There were 36 severe adverse events related to nimotuzumab; 16.7 % of them showed a definitive causal effect, 25 % were probable and 58.3 % were possible (Sierra P, 2012, personal communication). Those included within the adverse events having an incidence of 5 % or more were: vomiting (16.7 %), nausea (11.1 %), gastrointestinal hemorrhage (5.6 %); or an allergic/immunological cause such as infusion reaction (8.3 %) and anaphylaxis (5.6 %).

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Antibody profiles in patients treated with tumor necrosis factor-alpha antagonists: new findings

Antibody profiles in patients treated with tumor necrosis factor-alpha antagonists: new findings

Anti-TNF-α agents, such as infliximab and etanercept, have been reported to be beneficial for RA patients not responsive to the conventional treatment [1-3,5]. Our study confirms that adalimumab, a new fully human anti-TNF-α monoclonal antibody, is also effective in improving the clinical scores in RA patients. Reduction of RF and anti-CCP antibody titres has been recently correlated with clinical improvement after infliximab therapy in RA patients [6-8]. It has been suggested that TNF-α blocking might display an inhibitory effect on the production of antibodies closely related with RA disease activity [6]. Actually, besides their diagnostic value, high RF serum levels were shown to be an independent predictor for deteriorating radiological damage, and a greater prevalence of anti-CCP antibodies was found in patients who develop severe radiological damage [31-36]. However, while RF titre reduction was also reported by other groups after infliximab and etanercept therapy [7,8,10,11,37-39] contrasting results were found regarding anti-CCP antibody levels [7,8,10,11,37-39]. Such a discrepancy might be, at least in part, related to the different periods of follow-up and to the modalities to measure the antibody levels in the different studies. In fact, most of the enrolled patients displayed an aggressive form of the disease with high anti- CCP antibody titres; but, not all the studies carried out serial serum sample dilutions and tried to reduce the batch to batch variability in performing the solid phase assays for anti-CCP detection. This could have made the laboratory tests not sensitive enough to detect variations in the antibody titres during treatment.

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Aplicacin del Anlisis de Componentes Principales en el proceso de fermentacin de un anticuerpo monoclonal

Aplicacin del Anlisis de Componentes Principales en el proceso de fermentacin de un anticuerpo monoclonal

In the Center of Molecular Immunology (Havana, Cuba) an effective therapeutic monoclonal antibody against head and neck cancer is produced. Given the great variability of the concentration of this antibody in the industrial fermentation stage of the plant, it became necessary to apply a multivariate analysis technique such as the Principal Component Analysis, in order to reduce data dimensionality and to explain the main sources of variability of the process. In order to carry out the Principal Component Analysis through the software THE UNSCRAMBLER, the determination of the critical parameters of the fermentation stage through a risk model based on input and output matrix using data from the campaign of the year 2014 was carried out. As a result, two main components were able to explain more than 99% of the total variance, and it was possible to defi ne the critical parameters that have the greatest contribution to the variability of the fermentation process. These results corroborated the practical experiences of specialists of the plant and allowed to give recommendations to consider in the Plan of Continuous Verifi cation of the Process as proposing the inclusion in the strategy of control of the process the variables temperature, the speed of agitation, dissolved oxygen and the culture duration.

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Protocolos abcam

Protocolos abcam

In general, the species of the host animal in which an antibody was raised is important when using a conjugated secondary antibody to detect an unconjugated primary. For immunohistochemistry, the primary antibody should be raised in a species as phylogenetically different as possible from the species of the sample. This is to avoid potential cross-reactivity of the secondary anti-immunoglobulin antibody with endogenous immunoglobulins in the sample. For instance, a primary antibody used to detect a protein in a sample from a mouse should not be raised in mouse or rat. A primary antibody raised in rabbit will be a more appropriate choice, followed by an anti-rabbit IgG secondary antibody conjugated to a detection molecule (enzyme, fluorochrome, biotin, etc.). This issue can be avoided if a conjugated primary antibody is available. For other techniques using samples that do not contain endogenous immunoglobulin, the choice of host species is less critical. An example would be western blotting of a cell lysate that is not expected to contain IgG. However, tissue lysates and tissue culture supernatants that contain serum will contain immunoglobulins. IgG will appear in western blots of reduced, denatured samples as bands at 50 and 25 kDa corresponding to the heavy and light chains of the IgG molecule.

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Use of silver nanoparticles increased inhibition of cell associated HIV 1 infection by neutralizing antibodies developed against HIV 1 envelope proteins

Use of silver nanoparticles increased inhibition of cell associated HIV 1 infection by neutralizing antibodies developed against HIV 1 envelope proteins

The discovery of an HIV-1 vaccine that elicits broadly efficient neutralizing antibodies still remains an elusive goal especially after the recent failure of the leading T cell based HIV vaccine in human efficacy trials [7]. The envelope glycoproteins gp120 and gp41 that are the main targets for neutralizing antibodies are partially shielded by N-linkedglycans and other structurally- imposed steric constraints that limit antibody access to potential neutralization epitopes. The complex level of antigenic diversity of HIV-1, the shielding of key epi- topes within the three dimensional structure of the native Env trimer, and the failure of newer versions of Env proteins to elicit broadly reactive antibodies have led to some pessimism regarding the potential to ever elicit high titers of neutralizing antibodies against diverse strains of HIV-1. Therefore there is a need to maximize the efficiency of whatever titers of neutralizing antibodies generated by vaccines [8].

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Prevalence and risk factors of isolated anti HBc antibody and occult hepatitis B infection in hemodialysis patients: a nationwide study

Prevalence and risk factors of isolated anti HBc antibody and occult hepatitis B infection in hemodialysis patients: a nationwide study

elements in the blood prior to the HD session. All samples were tested for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels by a colorimetric method. In addition to checking HBsAg and anti-HBs (Both by Hepanosticka Biome- rieux, Boxtel, The Netherlands), the patients were also screened for human immunodeficiency virus (HIV) 1 and 2 (ELISA, MP Biomedicals, Illkirch, France), and anti-HBc by enzyme-linked immuno- sorbent assays (ELISA, Abbott Laboratories, US) every three months. ELISA generation III is the as- say that is usually used to check for HCV antibody. Qualitative HBV DNA (Roche Diagnostics GmbH, Mannheim, Germany) was checked for cases with positive HBsAg or isolated anti-HBc. HCV RNA was ordered for all cases with positive anti-HCV (Biorad, Segrate, Italy).

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Radioactive immunocomplex formation in a double antibody ria

Radioactive immunocomplex formation in a double antibody ria

Tube series were prepared with 100 µL of each of the different labelled and unlabelled antigen solutions, together with 100 µL of antibody solution and a bead in each tube. They were left to react in agitation for different time periods, after which they were washed, eliminating the liquid and leaving the bead in order to measure its radioactivity on the counter. One tube from each series was left to react for 24 hours, this being considered infinite time and therefore corresponding to the value at equilibrium.

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Gammapatía monoclonal: mieloma múltiple IgD. Reporte de un caso

Gammapatía monoclonal: mieloma múltiple IgD. Reporte de un caso

La enfermedad se estadifica estimando la masa tumoral mielomatosa en base a la cantidad de proteína monoclonal presente en el suero y/u orina (proteína M), con asociación con otros parámetros clínicos como la hemoglobina, las concentraciones de calcio sérico, el número de lesiones óseas líticas y la presencia o ausencia de insuficiencia renal (1) .

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Gamopatía monoclonal de significado incierto y otras entidades asociadas a paraproteínas monoclonales

Gamopatía monoclonal de significado incierto y otras entidades asociadas a paraproteínas monoclonales

Se suma el riesgo de trasformación maligna, una serie de condiciones asociadas a efectos patológicos inducidos por la paraproteína monoclonal. Este es un creciente grupo de enfermedades renales, neurológicas, dermatológicas y trastornos multisistémicos (5). En la mayoría de estas condiciones la población maligna es mínima y con frecuencia la gamopatía monoclonal subyacente no es sospechada. De importancia clínica es que el tratamiento antineoplásico puede lograr la regresión de algunos de estos cuadros.

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Coproantgeno Monoclonal para Deteccin de Helicobacter pylori en Nios  Evaluacin Inicial

Coproantgeno Monoclonal para Deteccin de Helicobacter pylori en Nios Evaluacin Inicial

Desde luego que por los pr opósitos de esta evaluación inicial con la pr ueba de copr oan tígeno monoclonal; no es posible por ahora hacer inferencias respecto al comportamiento de este padecimiento, o la sensibilidad o especificidad de la misma; esto podrá ser permisible al ten er un mayor númer o de pacientes estudiados, toda vez que hemos conocido el procedimiento y apreciado las bondades inmediatas del método 1,2,5,9,12 .

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Gammapatía monoclonal de significado incierto: A propósito de un caso de nefropatía por depósito de cadenas ligeras lambda

Gammapatía monoclonal de significado incierto: A propósito de un caso de nefropatía por depósito de cadenas ligeras lambda

Nephropathy associated with monoclonal gammopathies is mainly due to light chain deposition. The paraproteinemic kidney diseases are lesions associated with deposition of intact immunoglobulins or fragments of immunoglobulins (heavy and light chains). The disease due to deposition of light chains is a rare condition characterized by deposition of monoclonal light chains in many organs and as for the kidney, predominantly in glomeruli and tubular basement membranes. The disease is frequently associated with lymphoproliferative disorders and the majority of cases are caused by deposition of kappa light chains. Although presented primarily in clinical pictures of malignancy, sometimes no hematological pathology is detected and is called idiopathic or “primary”. It usually manifests as severe renal failure with nephrotic proteinuria, has not a clearly established treatment and the prog- nosis is poor. The clinical and histological features of the second case reported in Colombia of a light chain deposition nephropathy diagnosed in the context of a kidney paraproteinemic disease without malignancy data, is presented. (Acta Med Colomb 2014; 39: 196-201).

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Mycoplasma pneumonia infections in school children of a tropical community

Mycoplasma pneumonia infections in school children of a tropical community

In previous studies Foy et al. ( 1 9 66 ) found t hat children 5-14 years old were the most common primary cases of infection in me family, acting as a reservoir and becoming the most vector. In contrast to mis observation, they considered mat the " cornmunicability .. at school was low compared with me transmission within the family (Foy et al. 1971). In the present study, the antibody prevalence of children who to Palmares Centro School, from an outside district was compared with me antibody prevalen ce of the children attending their own district's school (Fig. 1). The risk of acquiring me infection was five times greater for children traveling to study in Palmares Centro, compared with the children studying in their own school district. The importance of the school in the transmission of me M. pneumoniae was a1so evident when children living' in a high prevalence district traveled to study in schools of lower prevalence. In this case, the risk of acquiring the infection was lower for the children who traveled (TabIe 2, Fig lB). Furthermore, there were no significant differences between the prevalence of M. pneumoniae in families with 1, 2, 3 or 4 siblings (TabIe 3). ,

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Gammapatía monoclonal: un diagnóstico a tener en cuenta

Gammapatía monoclonal: un diagnóstico a tener en cuenta

La exploración más sistemática de los pacientes y la mejora de los métodos electroforéticos en términos de normalización y sensibilidad, han hecho que el hallazgo fortuito de una gammapatía monoc lonal sea cada vez más frecuente y sin relación alguna con un contexto clínico que la sugiera . De esta forma, del total de las GM, cerca del 60% corresponde a una gammapatía monoclonal esencial o de significado incierto (GMSI) la cual, a pesar de ser asint omática y no requerir terapia, puede evolucionar a una GM maligna, lo que le da una condición de pre -maligna, con un riesgo de progresión del 1% al año. 3,4

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Fractura vertebral en una paciente con gammapatía monoclonal y neuropatía periférica

Fractura vertebral en una paciente con gammapatía monoclonal y neuropatía periférica

La fisiopatología de la NP asociada a MGUS no es del todo bien conocida. Los estudios anatomo- patológicos de pacientes con MW y NP asociada a MGUS IgM identificaron desmielinización y en- sanchamiento de la vaina de mielina. Se han podi- do demostrar depósitos de IgM monoclonal en esos engrosamientos de la mielina y en los detritus de tejido neuronal dentro de macrófagos y células de Schwann. Un estudio también encontró desmielini- zación en 5 pacientes con NP asociada a MGUS IgG y en 3 formación de “catáfilas de cebolla” (10) .

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2017.02.02.5

2017.02.02.5

  Protein A and protein G are bacterial proteins from Sta- phylococcus aureus and Streptococcus respectively, which natu- rally interacts with the Fcγ fragment of antibodies. This feature makes them extremely useful selective ligands, for several routine applications. Examples include the purification of monoclonal, polyclonal IgG-type antibodies and its subclasses, and the ad- sorption and purification of immune complexes involving IgG. IgG subclasses can be isolated from ascites, cell culture super- natants and serum 4, 21 . Another important factor to consider in

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