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1.
Bioprocess Biosyst Eng ; 47(7): 1039-1056, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38744689

RESUMEN

Cosmetics have been extremely popular throughout history and continue to be so today. Cosmetic and personal care products, including toothpaste, shampoo, lotions, and makeup, are typically made with petroleum-based surfactants. Currently, there is an increasing demand to enhance the sustainability of surface-active compounds in dermal formulations. Biosurfactants, derived from living cells, are considered more environmentally friendly than synthetic surfactants. Thus, the use of biosurfactants is a promising strategy for formulating more environmentally friendly and sustainable dermal products. Biosurfactants have the potential to replace chemical surface-active agents in the cosmetic sector due to their multifunctional qualities, such as foaming, emulsifying, and skin-moisturizing activities.In this study, two glycolipopeptide biosurfactants derived from Lactiplantibacillus plantarum OL5 were used as stabilizing factors in oil-in-water emulsions in the presence of coconut oils. Both biosurfactants increased emulsion stability, particularly in the 1:3 ratio, dispersion, and droplet size. Moreover, the cytotoxicity of the two Lactiplantibacillus plantarum biosurfactants was assessed on B lymphocytes and MCF-7 cells. Overall, the results gathered herein are very promising for the development of new green cosmetic formulations.


Asunto(s)
Cosméticos , Tensoactivos , Cosméticos/química , Tensoactivos/química , Tensoactivos/farmacología , Humanos , Células MCF-7 , Cuidados de la Piel , Emulsiones/química , Lactobacillus plantarum/metabolismo
2.
Cancer Immunol Immunother ; 71(5): 1139-1155, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-34608548

RESUMEN

BACKGROUND: CD155 immune checkpoint has recently emerged as a compelling immunotherapeutic target. Epigenetic DNA methylation changes are recognized as key molecular mechanisms in cancer development. Hence, the identification of methylation markers that are sensitive and specific for breast cancer may improve early detection and predict prognosis. We speculate that CD155 promoter methylation can be a valuable epigenetic biomarker, based upon strong indications for its immunoregulatory functions. METHODS: Methylation analyses were conducted on 14 CpGs sites in the CD155 promoter region by bisulfite pyrosequencing. To elucidate the related gene expression changes, a transcriptional study using RT-qPCR was performed. Statistical analyses were performed to evaluate correlations of CD155 methylation profiles with mRNA expression together with clinical-pathological features, prognosis and immune infiltrate. RESULTS: CD155 promoter methylation profile was significantly associated with SBR grade, tumor size, molecular subgroups, HER2 and hormonal receptors expression status. Low CD155 methylation rates correlated with better prognosis in univariate cox proportional hazard analysis and appeared as an independent survival predictor in cox-regression multivariate analysis. Further, methylation changes at CD155 specific CpG sites were consistent with CD155 membranous mRNA isoform expression status. Statistical analyses also showed a significant association with immune Natural Killer cell infiltrate when looking at the CpG7, CpG8, CpG9 and CpG11 sites. CONCLUSION: Altogether, our results contribute to a better understanding of the impact of CD155 immune checkpoint modality expression in breast tumors, revealing for the first time that specific CpG sites from CD155 promoter may be a potential biomarker in breast cancer monitoring.


Asunto(s)
Neoplasias de la Mama , Neoplasias de la Mama/metabolismo , Islas de CpG , Metilación de ADN , Epigénesis Genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Pronóstico , Regiones Promotoras Genéticas , Receptores Virales
3.
Sci Rep ; 11(1): 7550, 2021 04 06.
Artículo en Inglés | MEDLINE | ID: mdl-33824367

RESUMEN

B7-H6 and PD-L1 belong to the B7 family co-stimulatory molecules fine-tuning the immune response. The present work investigates the clinical effect of B7-H6 protein expression with PD-L1 status and the infiltration of natural killer cells as potential biomarkers in breast tumor inflammatory microenvironment. The expression levels of B7-H6 protein by cancer cells and immune infiltrating cells in human breast cancer tissues and evaluate their associations with PD-L1 expression, NK cell status, clinical pathological features and prognosis were explored. The immunohistochemistry labeling method was used to assess B7-H6 and PD-L1 proteins expression by cancer and immune cells. The associations between immune checkpoint, major clinical pathological variables and survival rates were analyzed. B7-H6 protein was depicted in both breast and immune cells. Results showed that Tumor B7-H6 expression is highly associated with Her-2 over expression. B7-H6 + immune cells are highly related to the Scarff-Bloom-Richardson grade and associated with PD-L1 expression and NK cells status. Survival analysis revealed a better prognosis in patients with low expression of B7-H6 by cancer cells. Conversely, B7-H6 + immune cells were significantly associated with longer survival. Findings strongly suggest an interaction between B7 molecules that contributes to a particular design of the inflammatory microenvironment. This may influence the efficiency of therapies based on antibodies blocking the PD-L1/PD1 pathway and can explain the detection of clinical benefits only in a fraction of patients treated with immune checkpoint inhibitors.


Asunto(s)
Antígenos B7/inmunología , Antígeno B7-H1/inmunología , Neoplasias de la Mama/inmunología , Microambiente Tumoral/inmunología , Adulto , Antígenos B7/análisis , Antígenos B7/metabolismo , Antígeno B7-H1/análisis , Biomarcadores de Tumor/inmunología , Mama/metabolismo , Mama/patología , Línea Celular Tumoral , Femenino , Humanos , Proteínas de Punto de Control Inmunitario/inmunología , Inmunohistoquímica/métodos , Células Asesinas Naturales/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Persona de Mediana Edad , Pronóstico
4.
Clin Breast Cancer ; 21(6): e738-e747, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-33727019

RESUMEN

BACKGROUND: Each subgroup of immune cells has a different prognostic role in breast cancer; however, the prognostic impact of tumor-infiltrating natural killer cells (TINKs) is still not well established. Our aim was to assess the prognostic impact of natural killer (NK) cells in breast carcinomas. MATERIALS AND METHODS: NK cells infiltration were assessed by immunohistochemistry (IHC). Statistical analyses were performed to evaluate the correlation of NK cells with clinical-pathological features and outcome. RESULTS: CD56 IHC was realized in 126 patients. NK cells infiltration showed significant and positive association with tumor high Scarff-Bloom-Richardson (SBR) grade. NK cells were significantly associated with HER2-positive breast cancer and triple-negative breast cancer subtypes. Analyses showed significant and inverse correlation with progesterone and estrogen receptors expression status. High NK cells were significantly related to high Ki-67 labeling index. Our data showed that high NK cells infiltrate was significantly associated with tumor-infiltrating lymphocytes in breast cancer tissues. At a median follow-up of 5.5 years, high CD56 expression (≥ 5 cells/10 high power field) was associated significantly with a good overall survival and with good disease-free survival. CONCLUSION: In this study, we assessed the important prognostic role of TINKs in breast carcinomas, which seems to be evident despite its association with aggressive pathological features. Thus evaluation of NK cells can be standardized and integrated in daily routine.


Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/inmunología , Células Asesinas Naturales/metabolismo , Linfocitos Infiltrantes de Tumor/inmunología , Adulto , Neoplasias de la Mama/patología , Supervivencia sin Enfermedad , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Pronóstico
5.
Life Sci ; 231: 116543, 2019 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-31176775

RESUMEN

AIMS: CD155 is a ligand of the NK activating receptor DNAM-1, it has been described in a variety of human malignancies, but its expression in breast cancer remains unclear and poorly studied. MAIN METHODS: CD155 expression and NK cells infiltration were investigated in 158 patients with breast cancer by immunohistochemistry (IHC). Statistical analyses were performed to evaluate correlations of CD155 expression with clinical-pathological features, prognosis and tumor immunity. KEY FINDINGS: Tumor cytoplasmic CD155 (cyt-CD155) was associated with lymphovascular invasion (p = 0.011), and membranous CD155 (m-CD155) was strongly correlated with the presence of Tumor Infiltrating natural killer cells (NK-TILs) (p = 0.0003). Survival analysis demonstrated that patients with high cyt-CD155 had a significantly worse overall survival (p < 0.001) and death free survival (p = 0.014) than those with low expression, while high levels of m-CD155 correlated with a better prognosis (p = 0.037). Furthermore, we found that patients with m-CD155Low/NKLow tumors had a significantly reduced overall survival (p = 0.012). Multivariate analysis showed that positive tumor m-CD155 status was a significant independent marker of good prognosis. Meanwhile, high cyt-CD155 expression was identified as an independent poor prognostic predictor, suggesting a key role in this malignancy. SIGNIFICANCE: Altogether, our results revealed that cyt-CD155 was associated with invasiveness and poorer prognosis, but the concomitant presence of m-CD155 and NK-TILs had an opposite prognostic relevance in breast cancer. These results raised the importance of CD155 IHC analysis to elucidate biomarker localization, leading to better understand and design therapeutic molecule targeting CD155 in breast tumors.


Asunto(s)
Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Receptores Virales/genética , Receptores Virales/aislamiento & purificación , Receptores Virales/metabolismo , Adulto , Anciano , Antígenos de Diferenciación de Linfocitos T/metabolismo , Neoplasias de la Mama/inmunología , Citoplasma/metabolismo , Supervivencia sin Enfermedad , Femenino , Humanos , Inmunohistoquímica , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Proteínas de la Membrana/metabolismo , Persona de Mediana Edad , Pronóstico , Receptores Virales/inmunología , Estudios Retrospectivos , Análisis de Supervivencia
6.
Toxicology ; 416: 44-53, 2019 03 15.
Artículo en Inglés | MEDLINE | ID: mdl-30721722

RESUMEN

Imipenem is a beta-Lactam antibiotic characterized by a broad spectrum of activity. It is prescribed to treat severe infections. Our goal is to investigate toxicity induced in male rat reproductive systems following exposure to this drug (15, 50 or 100 mg/kg) compared to gentamicin (50 mg/kg) treatment. Effects of imipenem on reproductive organ weights, histoarchitecture, sperm parameters, and oxidative stress parameters were evaluated. Serum testosterone levels were measured. Apoptosis and inflammatory behaviors were investigated by immunohistochemical proteins expression analysis of apoptosis regulator BAX (Bax), B-cell lymphoma 2 (Bcl-2), and interleukin-1 beta (IL-1 beta) in testis. Results showed a significant decrease in male fertility parameters including sperm count, sperm motility, reproductive organ weights and serum testosterone levels after imipenem administration as compared to the control and gentamicin treated groups. Increased sperm abnormality was significant in animals treated with high doses of imipenem. Oxidative stress analysis revealed an expressed increase in lipid peroxidation and carbonyl groups levels in testicular tissues compared to control. Similar results were observed with superoxide dismutase and catalase activities from testicular tissues. In addition, severe testicular lesions were observed in the seminiferous tubules as well as important impairments in spermatogenesis testifying an inflammatory microenvironment confirmed by the intensive expression of IL1-beta and Bax protein by germinal cells and Bcl-2 by Leydig cells. In conclusion, imipenem treatment with high doses was found to lead to oxidative stress in male reproductive organs and an inflammatory microenvironment leading to spermatogenesis dysfunction and histopathological changes in the testis.


Asunto(s)
Antibacterianos/toxicidad , Microambiente Celular , Imipenem/toxicidad , Infertilidad Masculina/inducido químicamente , Mediadores de Inflamación/metabolismo , Estrés Oxidativo/efectos de los fármacos , Testículo/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Epidídimo/efectos de los fármacos , Epidídimo/metabolismo , Epidídimo/patología , Gentamicinas/toxicidad , Infertilidad Masculina/metabolismo , Infertilidad Masculina/patología , Peroxidación de Lípido/efectos de los fármacos , Masculino , Carbonilación Proteica/efectos de los fármacos , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Espermatogénesis/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Espermatozoides/metabolismo , Espermatozoides/patología , Testículo/metabolismo , Testículo/patología , Testosterona/sangre
7.
Toxicol Mech Methods ; 29(5): 355-367, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-30621503

RESUMEN

This study aimed to evaluate the protective effect of a polysaccharide extracted from fenugreek seeds (Trigonella foenum-graecum) (FWEP) against insecticide-thiamethoxam (TMX)-induced hepatotoxicity. Obtained data exhibited potent antioxidant and antibacterial potentialities. On other trend, in vivo, adult female rats were divided into four groups: controls; TMX (100 mg/kg of body weight); TMX + FWEP at two graded doses, respectively (100 and 200 mg/kg of body weight) for 30 d. Up to TMX treatment, our data showed a significant increase in plasma markers of hepatotoxicity including alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and in lactate dehydrogenase (LDH) activities, which is coordinated with decline in total protein and albumin levels. Remarkably, a clear sign of genotoxicity was delivered by total disruption in hematological parameters and micronucleus (MN) test shown by severe chromatin degradation. These data were also associated with oxidative stress set up, histological and DNA injuries. However, co-administration with FWEP succeeded significantly in a dose-dependent manner in reducing and healing liver's hematological and genotoxic induced by TMX injuries.


Asunto(s)
Antioxidantes/uso terapéutico , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Daño del ADN/efectos de los fármacos , Contaminantes Ambientales/toxicidad , Polisacáridos/uso terapéutico , Tiametoxam/toxicidad , Trigonella/química , Animales , Antioxidantes/aislamiento & purificación , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Extractos Vegetales/química , Polisacáridos/aislamiento & purificación , Ratas Wistar , Semillas/química
8.
Int J Biol Macromol ; 123: 1267-1277, 2019 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-30521902

RESUMEN

The present study aims to investigate the physicochemical and the structural features of polysaccharide isolated from the red marine macro alga Chondrus canaliculatus (C.C.P) using FT-IR, gel filtration high-pressure chromatograph, HPLC-FID and solid state 13C NMR analysis. C.C.P was even more tested in vitro for its potential antioxidant properties and in vivo for its hemato-nephroprotective effects against fungicide - maneb (MB) - induced toxicity. Animals treated for 20 days were allocated into six groups per six rats each: group 1 served as vehicle control, group 2 received MB, group 3 received MB + C.C.P (100 mg/kg), group 4 MB + C.C.P (200 mg/kg), group 5 and 6 used as a positive control groups receiving only C.C.P; one 100 and the other 200 mg/kg, respectively. After MB injection, our data displayed a significant disruption in all hematological parameters associated with clear signs of nephrotoxicity. However, co-treatment with C.C.P at two graded doses led to an effective healing process against MB's hematological, biochemical, and histological kidney's oxidative injuries. In summary, our data suggest that C.C.P could be a prospective potent antioxidant, nephro and hemato-protective agent.


Asunto(s)
Antioxidantes/farmacología , Hematología , Riñón/patología , Maneb/toxicidad , Polisacáridos/química , Polisacáridos/farmacología , Sustancias Protectoras/farmacología , Algas Marinas/química , Animales , Compuestos de Bifenilo/química , Recuento de Células Sanguíneas , Proteína C-Reactiva/metabolismo , Espectroscopía de Resonancia Magnética con Carbono-13 , Quelantes/farmacología , Femenino , Depuradores de Radicales Libres/farmacología , Riñón/efectos de los fármacos , Recuento de Leucocitos , Malondialdehído/metabolismo , Metales/aislamiento & purificación , Peso Molecular , Monosacáridos/análisis , Picratos/química , Polisacáridos/aislamiento & purificación , Ratas Wistar , Espectroscopía Infrarroja por Transformada de Fourier , beta Caroteno/metabolismo
9.
Biol Trace Elem Res ; 184(1): 99-113, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28940108

RESUMEN

The current study aimed at evaluating the ability of a mineral and antioxidant-rich extract from Chondrus canaliculatus to improve maneb (MB)-induced toxicity in adult rat. The animals were divided into four groups: group 1 used as a control group, group 2 received MB, group 3 received MB + C. canaliculatus extract, and group 4 received only the algal extract. MB, a Mn-containing ethylene-bis-dithiocarbamate fungicide, induced oxidative stress damages, mineral perturbations in the plasma, urine, and bone, and genotoxicity in rats. Hematological analysis revealed in the MB-treated group a disruption in the number of red blood cells, platelets, and white blood cells associated with a striking genotoxicity. Interestingly, a significant increase in malondialdehyde and advanced oxidation protein product levels in erythrocytes and bones were found. On the other hand, an impairment of the antioxidant status in both tissues was occurred. Along, our results revealed that MB injection caused a striking drop and disruption in bone's mineral rates, especially calcium and phosphorus. These biochemical results were in accordance with the histological and molecular changes. However, co-treatment with C. canaliculatus extract showed, for the first time, that this alga was effective against MB-induced hematotoxicity, genotoxicity, and oxidative stress in the blood and bone and maintained osteomineral metabolism and bone histo-architecture. Such observations might be explained by the strong in vitro antioxidant and antibacterial activities exhibited by the alga, as well as by its high levels in several minerals: calcium, phosphorus, sodium, potassium, magnesium, iron, and zinc.


Asunto(s)
Antioxidantes/química , Antioxidantes/farmacología , Chondrus/química , Maneb/farmacología , Animales , Huesos/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Eritrocitos/efectos de los fármacos , Femenino , Malondialdehído/metabolismo , Oxidación-Reducción/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar
10.
Arch Physiol Biochem ; 124(4): 313-325, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-29171301

RESUMEN

CONTEXT: Hypercholesterolemia has significant cardiac consequences, since it is among the major risk factors of ischemic heart diseases. OBJECTIVE: The aim was searching the cardioprotective effect of chemical constituents from the sea lettuce Ulva lactuca upon hypercholesterolemic regime in mice. MATERIAL AND METHODS: Mice were randomly divided into three groups: untreated group, hypercholesterolemic group, and mice receiving 1% cholesterol associated with U. lactuca ethanolic extract. RESULTS: In vitro study demonstrated that algal extract has antioxidant efficacy attributable to the presence of phenolic compounds. Additionally, the alga alleviated cardiotoxicity, as shown by the improvement of haematological parameters, white cell viability, heart oxidative stress, plasma biochemical parameters and index of atherogenesis. Gene expression of the proinflammatory cytokines TNF-α, IL-1ß and IL-6 significantly decreased in the heart of U. lactuca supplemented hypercholesterolemic animals. CONCLUSION: It was established that the green alga, thanks to its bioactive compounds, effectively counteracts cardiotoxic effects of hypercholesterolemic regime.


Asunto(s)
Anticolesterolemiantes/uso terapéutico , Productos Biológicos/uso terapéutico , Cardiotónicos/uso terapéutico , Suplementos Dietéticos , Hipercolesterolemia/prevención & control , Algas Marinas/química , Ulva/química , Animales , Anticolesterolemiantes/química , Antioxidantes/uso terapéutico , Aterosclerosis/etiología , Aterosclerosis/prevención & control , Biomarcadores/sangre , Biomarcadores/metabolismo , Cardiotónicos/química , Enfermedad Coronaria/etiología , Enfermedad Coronaria/prevención & control , Citocinas/genética , Citocinas/metabolismo , Suplementos Dietéticos/análisis , Etanol/química , Regulación de la Expresión Génica , Hipercolesterolemia/metabolismo , Hipercolesterolemia/patología , Hipercolesterolemia/fisiopatología , Ratones , Miocardio/inmunología , Miocardio/metabolismo , Miocardio/patología , Estrés Oxidativo , Distribución Aleatoria , Solventes/química
11.
Toxicol Mech Methods ; 24(9): 679-87, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25179310

RESUMEN

Methylthiophanate is one of the widely used fungicides to control important fungal diseases of crops. The aim of this study was to elucidate the short-term hematoxicity and genotoxicity effects of methylthiophanate administered by intraperitoneal way at three doses (300, 500 and 700 mg/kg of body weight) after 24, 48 and 72 h. Our results showed, 24 h after methylthiophanate injection, a hematological perturbation such as red blood cells (p < 0.05, p < 0.05 and p < 0.01) and hemoglobin content (p < 0.05), respectively, and a noticeable genotoxic effect in WBC evidenced by a significant increase in the frequency of the micronuclei and a decrease in cell viability. An increase in erythrocyte osmotic fragility was also noted after 24 and 48 h of methylthiophanate treatment at graded doses. A significant increase in hydrogen peroxide, advanced oxidation of protein products and malondialdehyde levels, in erythrocytes of methylthiophanate-treated rats with 300, 500 and 700 mg/kg of body weight, was also observed after 24 h of treatment (p < 0.05, p < 0.01 and p < 0.001, respectively), suggesting the implication of oxidative stress in its toxicity. Antioxidants activities of superoxide dismutase and glutathione peroxidase in erythrocytes significantly increased (p < 0.001) 24 h after the highest dose injected. While all these parameters were improved after 72 h of methylthiophanate injection (300, 500 and 700 mg/kg body weight). In conclusion, these data showed that the exposure of adult rats to methylthiophanate resulted in oxidative stress leading to hematotoxicity and the impairment of defence system, confirming the pro-oxidant and genotoxic effects of this fungicide.


Asunto(s)
Especies Reactivas de Oxígeno/metabolismo , Tiofanato/farmacología , Animales , Dosificación Letal Mediana , Masculino , Ratas , Ratas Wistar , Tiofanato/química
12.
Viral Immunol ; 19(2): 324-34, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16817775

RESUMEN

Morbillivirus infections have been known for a long time to be associated with an acute immunosuppression in their natural hosts. Here, we show that recombinant Morbillivirus nucleoproteins from canine distemper virus, peste-des-petits-ruminants virus, and Rinderpest virus bind B-lymphocytes from dogs, goats, and cattle, respectively, similarly to measles virus nucleoprotein in humans. The use of surface plasmon resonance imaging allowed the real time detection of differential interactions between Morbillivirus nucleoproteins and FcgammaRIIb (CD32). Moreover, those nucleoproteins which bind murine Fcgamma receptor inhibited the inflammatory immune responses in mice in a Fc receptor- dependent manner. In contrast, nucleoprotein from closely related Henipavirus genus, belonging to the Paramyxoviridae family as Morbillivirus, was devoid of capacity either to bind FcgammaRIIb or to inhibit inflammatory response. Altogether, these results suggest that nucleoprotein-FcR interaction is a common mechanism used by different Morbilliviruses to modulate the immune response.


Asunto(s)
Terapia de Inmunosupresión , Infecciones por Morbillivirus/inmunología , Morbillivirus/patogenicidad , Nucleoproteínas/metabolismo , Animales , Baculoviridae/genética , Baculoviridae/metabolismo , Bovinos , Línea Celular , Perros , Humanos , Ratones , Ratones Endogámicos C57BL , Morbillivirus/clasificación , Morbillivirus/genética , Morbillivirus/inmunología , Infecciones por Morbillivirus/virología , Nucleoproteínas/genética , Receptores de IgG/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Resonancia por Plasmón de Superficie
13.
Clin Chem ; 52(2): 255-62, 2006 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-16339301

RESUMEN

BACKGROUND: Developing rapid, high-throughput assays for detecting and characterizing protein-protein interactions is a great challenge in the postgenomic era. We have developed a new method that allows parallel analysis of multiple analytes in biological fluids and is suitable for biological and medical studies. METHODS: This technology for studying peptide-antibody interactions is based on polypyrrole-peptide chips and surface plasmon resonance imaging (SPRi). We generated a chip bearing a large panel of peptide probes by successive electro-directed copolymerizations of pyrrole-peptide conjugates on a gold surface. RESULTS: We provide evidence that (a) the signal produced by antibody binding is highly specific; (b) the detected signal specifically reflects the antibody concentration of the tested solution in a dose-dependent manner; (c) this technique is appropriate for analyzing complex media such as undiluted sera, a novelty with respect to previous techniques; and (d) correlation between classic ELISA results and the SPRi signal is good (P = 0.008). We also validated this system in a medical model by detecting anti-hepatitis C antibodies in patient-derived sera. CONCLUSION: Because of its characteristics (easy preparation of the peptide chip; high-throughput, label-free, real-time detection; high specificity; and low background), this technology is suitable for screening biological samples and for large-scale studies.


Asunto(s)
Sitios de Unión de Anticuerpos , Anticuerpos contra la Hepatitis C/química , Péptidos/química , Polímeros/química , Análisis por Matrices de Proteínas/métodos , Pirroles/química , Resonancia por Plasmón de Superficie , Química Clínica , Oro , Hepacivirus/inmunología , Humanos , Análisis por Matrices de Proteínas/instrumentación , Mapeo de Interacción de Proteínas , Proteínas Recombinantes/química , Sensibilidad y Especificidad , Albúmina Sérica Bovina/química
14.
Anal Biochem ; 347(2): 193-200, 2005 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-16266681

RESUMEN

Protein microarray is a promising technology that should combine rapidity and easy use with high throughput and versatility. This article describes a method in which an electrocopolymerization process is employed to graft biological molecules on to a chip so that surface plasmon resonance imaging may be used to detect molecular interactions. Copolymerization of pyrrole-modified protein and pyrrole is an efficient grafting process which immobilizes molecules at defined positions on a gold surface. Surface plasmon resonance imaging is an optical technique that allows real-time simultaneous detection of molecular interactions on a large number of spots without labeling. This method was successfully used to analyze antibody-antigen interactions. This illustrates its high specificity and good sensitivity and demonstrates its suitability for biological studies.


Asunto(s)
Reacciones Antígeno-Anticuerpo , Análisis por Matrices de Proteínas/métodos , Animales , Técnicas de Química Analítica , Gonadotropina Coriónica/inmunología , Humanos , Técnicas In Vitro , Muramidasa/inmunología , Polímeros , Análisis por Matrices de Proteínas/instrumentación , Pirroles , Resonancia por Plasmón de Superficie , Propiedades de Superficie
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