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1.
Science ; 372(6537)2021 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-33795432

RESUMEN

Multivalent display of receptor-engaging antibodies or ligands can enhance their activity. Instead of achieving multivalency by attachment to preexisting scaffolds, here we unite form and function by the computational design of nanocages in which one structural component is an antibody or Fc-ligand fusion and the second is a designed antibody-binding homo-oligomer that drives nanocage assembly. Structures of eight nanocages determined by electron microscopy spanning dihedral, tetrahedral, octahedral, and icosahedral architectures with 2, 6, 12, and 30 antibodies per nanocage, respectively, closely match the corresponding computational models. Antibody nanocages targeting cell surface receptors enhance signaling compared with free antibodies or Fc-fusions in death receptor 5 (DR5)-mediated apoptosis, angiopoietin-1 receptor (Tie2)-mediated angiogenesis, CD40 activation, and T cell proliferation. Nanocage assembly also increases severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus neutralization by α-SARS-CoV-2 monoclonal antibodies and Fc-angiotensin-converting enzyme 2 (ACE2) fusion proteins.


Asunto(s)
Anticuerpos/química , Anticuerpos/inmunología , Nanoestructuras , Ingeniería de Proteínas , Transducción de Señal , Angiopoyetinas/química , Angiopoyetinas/inmunología , Angiopoyetinas/metabolismo , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/inmunología , Anticuerpos Neutralizantes/química , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/química , Anticuerpos Antivirales/inmunología , Linfocitos B/inmunología , Antígenos CD40/química , Antígenos CD40/inmunología , Antígenos CD40/metabolismo , Línea Celular Tumoral , Proliferación Celular , Simulación por Computador , Genes Sintéticos , Humanos , Fragmentos Fc de Inmunoglobulinas/química , Activación de Linfocitos , Modelos Moleculares , Unión Proteica , Receptor TIE-2/metabolismo , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/inmunología , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , Linfocitos T/inmunología , Linfocitos T/fisiología
2.
Methods Mol Biol ; 2265: 223-233, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704718

RESUMEN

The advent of personalized medicines targeting cell signaling pathways has radically improved melanoma patient outcomes. More recently, immune-modulating therapies disrupting the PD-1/PD-L1 axis have become a powerful tool in the treatment of a range of melanoma, showing a profound improvement in the overall survival outcomes. However, immune checkpoint inhibitors (ICIs) are associated with considerable toxicities and appear to only be efficacious in a subset of melanoma patients. Therefore, there is an urgent need to identify biomarkers that can determine if patients will or will not respond to ICI therapy. Here, we describe an optimized method for analyzing PD-L1 expression on circulating melanoma cells following immunomagnetic enrichment from patient blood samples.


Asunto(s)
Antígeno B7-H1/metabolismo , Separación Inmunomagnética/métodos , Melanoma/sangre , Células Neoplásicas Circulantes/inmunología , Anticuerpos/inmunología , Antígeno B7-H1/inmunología , Humanos , Leucocitos Mononucleares/citología , Biopsia Líquida/métodos , Melanoma/diagnóstico
3.
Food Chem ; 345: 128812, 2021 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-33601655

RESUMEN

Due to complex matrixes and specific reagent deficiency, the rapid detection of histamine is still a challenge to date. Based on the high peroxidase-like activity of iron-cobalt co-doped carbon dots, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established for histamine detection using the mimic enzyme labeled with histamine antibody (His-Ab). Through the competitive binding of the labeled His-Ab to solid-phase and sample antigens, histamine content was detected with a linear range of 2.5-150 µg mL-1. The detection limit based on 3σ/K was 0.50 mg kg-1, which was much lower than those of commercial His-kit and HPLC methods. The ic-ELISA method was applied to histamine detection in fish samples with the recovery of (103.4 ± 0.5)%, which was in accord with those of commercial His-kit and HPLC methods. The results indicated that the established ic-ELISA method was suitable for rapid detection of histamine in fish samples with high accuracy, sensitivity and stability.


Asunto(s)
Peces/metabolismo , Histamina/análisis , Puntos Cuánticos/química , Animales , Anticuerpos/química , Anticuerpos/inmunología , Carbono/química , Cobalto/química , Ensayo de Inmunoadsorción Enzimática , Histamina/inmunología , Hierro/química , Límite de Detección , Reproducibilidad de los Resultados , Alimentos Marinos/análisis
4.
Food Chem ; 350: 129229, 2021 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-33636619

RESUMEN

A new strategy to mimic antibody for electrochemical recognition and detection of deoxynivalenol (DON) using a highly-sensitive and selective antibody-like sensor based on molecularly imprinted poly(l-arginine) (P-Arg-MIP) on carboxylic acid functionalized carbon nanotubes (COOH-MWCNTs) was proposed. l-arginine as functional monomer was screened to prepare imprinted electrode via its electro-polymerization in the presence of DON onto the surface of COOH-MWCNTs electrode coupled with theoretical calculation. Surface morphology, structural characteristics, and electrochemical properties of P-Arg-MIP/COOH-MWCNTs were characterized by SEM, EDS, FTIR, and CV, respectively. P-Arg-MIP/COOH-MWCNTs displayed relatively high conductivity, high effective surface area, antibody-like molecular recognition and affinity, and a good response towards DON in a linear range from 0.1 to 70 µM with LOD of 0.07 µM in wheat flour samples with satisfactory recovery and feasible practicability in comparison with HPLC. This method provides a promising biomimetic sensing platform for the determination of mycotoxins in food and agro-products.


Asunto(s)
Biomimética/instrumentación , Límite de Detección , Impresión Molecular , Nanotubos de Carbono/química , Péptidos/química , Péptidos/síntesis química , Tricotecenos/análisis , Anticuerpos/inmunología , Electroquímica , Electrodos , Harina/análisis , Tricotecenos/química , Triticum/química
5.
Cancer Sci ; 112(4): 1417-1428, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33539630

RESUMEN

Chimeric antigen receptor (CAR)-T cell therapy has shown salient efficacy in cancer immunotherapy, particularly in the treatment of B cell malignancies. However, the efficacy of CAR-T for solid tumors remains inadequate. In this study, we displayed that c-met is an appropriate therapeutic target for papillary renal cell carcinoma (PRCC) using clinical samples, developed an anti-human c-met CAR-T cells, and investigated the anti-tumor efficacy of the CAR-T cells using an orthotopic mouse model as pre-clinical research. Administration of the anti-c-met CAR-T cells induced marked infiltration of the CAR-T cells into the tumor tissue and unambiguous suppression of tumor growth. Furthermore, in combination with axitinib, the anti-tumor efficacy of the CAR-T cells was synergistically augmented. Taken together, our current study demonstrated the potential for clinical application of anti-c-met CAR-T cells in the treatment of patients with PRCC.


Asunto(s)
Carcinoma de Células Renales/inmunología , Carcinoma de Células Renales/terapia , Neoplasias Renales/inmunología , Neoplasias Renales/terapia , Proteínas Proto-Oncogénicas c-met/inmunología , Receptores Quiméricos de Antígenos/inmunología , Linfocitos T/inmunología , Anciano , Animales , Anticuerpos/inmunología , Antígenos de Neoplasias/inmunología , Línea Celular Tumoral , Células Cultivadas , Modelos Animales de Enfermedad , Femenino , Humanos , Inmunoterapia/métodos , Masculino , Ratones , Ratones Endogámicos NOD , Ensayos Antitumor por Modelo de Xenoinjerto/métodos
6.
J Vis Exp ; (168)2021 02 08.
Artículo en Inglés | MEDLINE | ID: mdl-33616097

RESUMEN

Measurements of the specificity and affinity of antigen-antibody interactions are critically important for medical and research applications. In this protocol, we describe the implementation of a new single-molecule technique, mass photometry (MP), for this purpose. MP is a label- and immobilization-free technique that detects and quantifies molecular masses and populations of antibodies and antigen-antibody complexes on a single-molecule level. MP analyzes the antigen-antibody sample within minutes, allowing for the precise determination of the binding affinity and simultaneously providing information on the stoichiometry and the oligomeric state of the proteins. This is a simple and straightforward technique that requires only picomole quantities of protein and no expensive consumables. The same procedure can be used to study protein-protein binding for proteins with a molecular mass larger than 50 kDa. For multivalent protein interactions, the affinities of multiple binding sites can be obtained in a single measurement. However, the single-molecule mode of measurement and the lack of labeling imposes some experimental limitations. This method gives the best results when applied to measurements of sub-micromolar interaction affinities, antigens with a molecular mass of 20 kDa or larger, and relatively pure protein samples. We also describe the procedure for performing the required fitting and calculation steps using basic data analysis software.


Asunto(s)
Afinidad de Anticuerpos , Complejo Antígeno-Anticuerpo/inmunología , Fotometría/métodos , Anticuerpos/inmunología , Humanos , Imagenología Tridimensional , Peso Molecular , Distribución Normal , Unión Proteica , Programas Informáticos , Trombina/inmunología
7.
Int J Mol Sci ; 22(2)2021 Jan 16.
Artículo en Inglés | MEDLINE | ID: mdl-33467096

RESUMEN

B cell activating factor (BAFF) is a cytokine that plays a role in the survival, proliferation and differentiation of B cells. We proposed to observe the effects of BAFF inhibition on the humoral immune responses of an allosensitized mouse model using HLA.A2 transgenic mice. Wild-type C57BL/6 mice were sensitized with skin allografts from C57BL/6-Tg (HLA-A2.1)1Enge/J mice and were treated with anti-BAFF monoclonal antibody (mAb) (named Sandy-2) or control IgG1 antibody. HLA.A2-specific IgG was reduced in BAFF-inhibited mice compared to the control group (Δ-13.62 vs. Δ27.07, p < 0.05). BAFF inhibition also resulted in increased pre-pro and immature B cell proportions and decreased mature B cells in the bone marrow (p < 0.05 vs. control). In the spleen, an increase in transitional B cells was observed with a significant decrease in marginal and follicular B cells (p < 0.05 vs. control). There was no significant difference in the proportions of long-lived plasma and memory B cells. Microarray analysis showed that 19 gene probes were significantly up- (>2-fold, p < 0.05) or down-regulated (≤2-fold, p < 0.05) in the BAFF-inhibited group. BAFF inhibition successfully reduced alloimmune responses through the reduction in alloantibody production and suppression of B cell differentiation and maturation. Our data suggest that BAFF suppression may serve as a useful target in desensitization therapy.


Asunto(s)
Factor Activador de Células B/antagonistas & inhibidores , Antígeno HLA-A2/inmunología , Inmunización , Aloinjertos/inmunología , Animales , Anticuerpos/inmunología , Linfocitos B/inmunología , Células de la Médula Ósea/inmunología , Inmunoglobulina G/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Trasplante de Piel/efectos adversos , Bazo/citología , Bazo/inmunología
8.
Nature ; 589(7843): 630-632, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33500572

Asunto(s)
Anticuerpos/uso terapéutico , Biología Celular , Biología Evolutiva , Nariz Electrónica , Espectrometría de Masas/instrumentación , Neurociencias , Animales , Anticuerpos/química , Anticuerpos/genética , Anticuerpos/inmunología , Proteínas Bacterianas/efectos de los fármacos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/efectos de la radiación , Bioimpresión/tendencias , /inmunología , /química , /provisión & distribución , Biología Celular/instrumentación , Biología Celular/tendencias , Biología Evolutiva/métodos , Biología Evolutiva/tendencias , Embrión de Mamíferos/citología , Embrión de Mamíferos/embriología , Embrión de Mamíferos/metabolismo , Desarrollo Embrionario/genética , Holografía/tendencias , Humanos , Inmunoglobulina E/química , Inmunoglobulina E/genética , Inmunoglobulina E/inmunología , Inmunoglobulina E/uso terapéutico , Canales Iónicos/metabolismo , Espectrometría de Masas/métodos , Proteínas de la Membrana/efectos de los fármacos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/efectos de la radiación , Ratones , Microscopía/instrumentación , Microscopía/tendencias , Sondas Moleculares/análisis , Neoplasias/tratamiento farmacológico , Neurociencias/métodos , Neurociencias/tendencias , Optogenética/tendencias , Análisis de la Célula Individual , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
9.
Nat Commun ; 12(1): 379, 2021 01 22.
Artículo en Inglés | MEDLINE | ID: mdl-33483508

RESUMEN

Allergic reactions occur when IgE molecules become crosslinked by antigens such as food proteins. Here we create the 'AllerScan' programmable phage display system to characterize the binding specificities of anti-allergen IgG and IgE antibodies in serum against thousands of allergenic proteins from hundreds of organisms at peptide resolution. Using AllerScan, we identify robust anti-wheat IgE reactivities in wheat allergic individuals but not in wheat-sensitized individuals. Meanwhile, a key wheat epitope in alpha purothionin elicits dominant IgE responses among allergic patients, and frequent IgG responses among sensitized and non-allergic patients. A double-blind, placebo-controlled trial shows that alpha purothionin reactivity, among others, is strongly modulated by oral immunotherapy in tolerized individuals. AllerScan may thus serve as a high-throughput platform for unbiased analysis of anti-allergen antibody specificities.


Asunto(s)
Alérgenos/inmunología , Anticuerpos/inmunología , Epítopos/inmunología , Biblioteca de Péptidos , Hipersensibilidad al Trigo/inmunología , Adolescente , Adulto , Alérgenos/genética , Anticuerpos/sangre , Péptidos Catiónicos Antimicrobianos/inmunología , Niño , Preescolar , Femenino , Humanos , Hipersensibilidad/inmunología , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Masculino , Persona de Mediana Edad , Proteínas de Plantas/inmunología , Adulto Joven
10.
Food Chem ; 336: 127718, 2021 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-32763741

RESUMEN

A novel dual near-infrared fluorescence-based lateral flow immunosensor was developed to determine zearalenone and deoxynivalenol in maize. Two near-infrared dyes with distinct fluorescence characteristics were utilized to separately label the anti-zearalenone and anti-deoxynivalenol antibodies as detection reagents. The capture antigens zearalenone-BSA and deoxynivalenol-BSA were mixed and immobilized on the same test line of nitrocellulose membrane. This assay format facilitates simultaneous detection of the two mycotoxins on a single test line. After optimizing experimental parameters, the limits of detection for zearalenone and deoxynivalenol were as low as 0.55 µg/kg and 3.8 µg/kg in maize, respectively. The spiking experiment yielded recovery ratios ranging from 81.7% to 107.3% with coefficients of variation less than 14% demonstrating high assay accuracy and precision. Moreover, the actual sample analysis produced consistent results between this method and instrumental method. Therefore, the developed immunosensor can serve as an accurate and efficient approach for monitoring mycotoxins in agricultural products.


Asunto(s)
Inmunoensayo/métodos , Tricotecenos/análisis , Zea mays/química , Zearalenona/análisis , Animales , Anticuerpos/inmunología , Bovinos , Colorantes Fluorescentes/química , Límite de Detección , Micotoxinas/análisis , Micotoxinas/inmunología , Reproducibilidad de los Resultados , Albúmina Sérica Bovina/química , Espectroscopía Infrarroja Corta , Tricotecenos/inmunología , Zea mays/metabolismo , Zearalenona/inmunología
11.
Methods Mol Biol ; 2237: 217-224, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33237421

RESUMEN

We describe here a standard protocol for determining the phosphorylation status of protein multiplexes using antibody arrays and a biotinylated Phos-tag with a dodeca(ethylene glycol) spacer (Phos-tag Biotin). The procedure is based on an antibody microarray technique used in conjunction with an enhanced chemiluminescence system, and it permits the simultaneous and highly sensitive detection of multiple phosphoproteins in a cell lysate. By using this procedure, we have demonstrated the quantitative detection of the entire phosphorylation status of a target protein involved in intracellular signaling.


Asunto(s)
Fosfoproteínas/análisis , Análisis por Matrices de Proteínas/métodos , Animales , Anticuerpos/inmunología , Biotinilación , Humanos , Inmunoensayo/métodos , Mediciones Luminiscentes/métodos , Fosfoproteínas/inmunología , Polietilenglicoles
12.
Methods Mol Biol ; 2237: 257-261, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33237425

RESUMEN

Antibody arrays have been widely applied in both basic research and clinical studies. Data analysis, archiving, and sharing of resulting data are very important for exploring and expanding the power of antibody microarray studies. The protein microarray database (PMD) has provided standards tailored for the management of protein microarray data and constructed an automated pipeline for array data analysis. In this chapter, we will describe the framework design, platform construction, and analysis tool integration of the PMD.


Asunto(s)
Bases de Datos de Proteínas , Análisis por Matrices de Proteínas/métodos , Programas Informáticos , Animales , Anticuerpos/inmunología , Humanos , Inmunoensayo/métodos
13.
Methods Mol Biol ; 2198: 217-226, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-32822035

RESUMEN

Immunostaining (also called as immunofluorescence) is a fluorescence labeling method to stain one or more epitopes of interest on DNA and/or protein using specific antibodies. Cytosine modifications can be detected quantitatively by immunostaining. The protocol commonly includes sequential steps. These include fixation, permeabilization, antigen retrieval, blocking, incubation with primary and secondary antibodies, and visualization under the microscope followed by image-based intensity analysis of staining. Each step is important, but antigen retrieval is especially necessary for DNA epitopes such as cytosine modifications as antibodies can access cytosines in DNA only once the DNA double-strand is denatured and DNA-packaging proteins have been removed. Hydrochloric acid is commonly used for this purpose. However, there are additional treatments with enzymes to enhance antigen retrieval and improve the detection by increasing staining intensity. This chapter describes current methodology for improving antigen retrieval for the staining of the cytosine modifications 5'-methylcytosine (5meC), 5'-hydroxymethylcytosine (5hmC), 5'-formylcytosine (5fC), and 5'-carboxycytosine (5caC).


Asunto(s)
Citosina/inmunología , Metilación de ADN/inmunología , Técnica del Anticuerpo Fluorescente/métodos , 5-Metilcitosina/análogos & derivados , 5-Metilcitosina/química , Animales , Anticuerpos/inmunología , Antígenos/metabolismo , Citosina/análogos & derivados , Citosina/química , Citosina/metabolismo , ADN/metabolismo , Metilación de ADN/genética , Epigénesis Genética/genética , Humanos
14.
Methods Mol Biol ; 2237: 1-10, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33237404

RESUMEN

Sandwich-based antibody arrays enable the detection of multiple proteins simultaneously, thus offering a time- and cost-effective alternative to single-plex platforms. The protein of interest is "sandwiched" between an antibody that captures it to the array and a second antibody that is used for detection. Here we describe a 1-day procedure to process samples, such as serum or cell lysates, with a quantitative sandwich-based antibody array on a glass substrate using fluorescence.


Asunto(s)
Técnica del Anticuerpo Fluorescente/métodos , Pruebas Inmunológicas/métodos , Análisis por Matrices de Proteínas/métodos , Animales , Anticuerpos/inmunología , Citocinas/análisis , Citocinas/inmunología , Humanos
15.
Methods Mol Biol ; 2237: 93-102, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33237411

RESUMEN

Antibody microarray is a fundamental, high-content technology for analyzing biomarkers with a multiplexity even at the proteomic level. Recent advancement in this field has driven the antibody array into a new territory related with single-cell analysis. Here we describe a flow pattern-based method for producing a high-density barcode antibody microarray for the detection of proteins in fluidic samples and in single cells. The antibody microarray is fabricated by a perpendicularly oriented flow patterning of single-stranded barcode DNAs, which are then converted into DNA-antibody conjugates. Compared to conventional microarrays, this barcode antibody microarray features a simple and high-throughput assay while achieving both high sensitivity and specificity. This barcode technology provides new clues for developing next-generation antibody microarrays and can be widely used in protein biomarker discovery, cell signaling network analysis, and disease diagnosis and prognosis.


Asunto(s)
Anticuerpos/química , ADN/química , Microfluídica/métodos , Análisis por Matrices de Proteínas/métodos , Anticuerpos/inmunología , Ensayos Analíticos de Alto Rendimiento/métodos , Inmunoensayo/métodos , Análisis de la Célula Individual/métodos
16.
Methods Mol Biol ; 2237: 103-122, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33237412

RESUMEN

Reverse phase protein arrays (RPPA) are used to quantify proteins and protein posttranslational modifications in cellular lysates and body fluids. RPPA technology is suitable for biomarker discovery, protein pathway profiling, functional phenotype analysis, and drug discovery mechanism of action. The principles of RPPA technology are (a) immobilizing protein-containing specimens on a coated slide in discrete spots, (b) antibody recognition of proteins, (c) amplification chemistries to detect the protein-antibody complex, and (d) quantifying spot intensity. Construction of a RPPA begins with the robotic liquid transfer of protein-containing specimens from microtiter plates onto nitrocellulose-coated slides. The robotic arrayer deposits each sample as discrete spots in an array format. Specimens, controls, and calibrators are printed on each array, thus providing a complete calibrated assay on a single slide. Each RPPA slide is subsequently probed with catalyzed signal amplification chemistries and a single primary antibody, a secondary antibody, and either fluorescent or colorimetric dyes. The focus of this chapter is to describe RPPA detection and imaging using a colorimetric (diaminobenzidine (DAB)) detection strategy.


Asunto(s)
Análisis por Matrices de Proteínas/métodos , 3,3'-Diaminobencidina/química , Animales , Anticuerpos/inmunología , Línea Celular , Colorimetría/métodos , Humanos , Inmunoensayo/métodos , Procesamiento Proteico-Postraduccional , Proteoma/inmunología , Proteoma/metabolismo
17.
Nat Rev Immunol ; 21(2): 83-100, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33353987

RESUMEN

Immunization is a cornerstone of public health policy and is demonstrably highly cost-effective when used to protect child health. Although it could be argued that immunology has not thus far contributed much to vaccine development, in that most of the vaccines we use today were developed and tested empirically, it is clear that there are major challenges ahead to develop new vaccines for difficult-to-target pathogens, for which we urgently need a better understanding of protective immunity. Moreover, recognition of the huge potential and challenges for vaccines to control disease outbreaks and protect the older population, together with the availability of an array of new technologies, make it the perfect time for immunologists to be involved in designing the next generation of powerful immunogens. This Review provides an introductory overview of vaccines, immunization and related issues and thereby aims to inform a broad scientific audience about the underlying immunological concepts.


Asunto(s)
Control de Enfermedades Transmisibles/métodos , Enfermedades Transmisibles/inmunología , Vacunas Atenuadas/inmunología , Vacunas de Productos Inactivados/inmunología , Vacunología/métodos , Anticuerpos/inmunología , Antígenos/inmunología , Humanos , Memoria Inmunológica/inmunología , Linfocitos T/inmunología , Vacunación
18.
Food Chem ; 339: 128084, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33152875

RESUMEN

Toxic small molecule contaminants (SMCs) residues in food threaten human health. Immunoassays are popular and simple techniques for SMCs analysis. However, immunoassays based on polyclonal antibodies, monoclonal antibodies and chemosynthetic antigens have some defects, such as complicated preparation of antibodies, risk of toxic haptens using for antigen chemosynthesis and so on. Phage-display technique has been proven to be an attractive alternative approach to producing antibody and antigen substitutes of SMCs, and opened up new realms for developing immunoassays of SMCs. These substitutes contain five types, including anti-idiotypic recombinant antibody (AIdA), anti-immune complex peptide (AIcP), anti-immune complex recombinant antibody (AIcA) and anti-SMC recombinant antibody (anti-SMC RAb). In this review, the principle of immunoassays based on the five types of substitutes, as well as their application and advantages are summarized and discussed.


Asunto(s)
Anticuerpos/inmunología , Antígenos/inmunología , Inmunoensayo/métodos , Biblioteca de Péptidos , Bibliotecas de Moléculas Pequeñas/análisis , Humanos
19.
Clin Nucl Med ; 46(3): 250-251, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33323741

RESUMEN

ABSTRACT: We report the case of a 55-year-old man presenting pseudopsychiatric behavior disorders of subacute-onset. MRI showed a FLAIR (fluid-attenuated inversion recovery) hyperintensity in the left hippocampus. The diagnosis of limbic encephalitis was raised, and the patient was referred for an 18F-FDG PET/CT. PET/CT depicted an increased uptake of the left mesiotemporal structures and also an increased uptake of both cerebellum and striatal areas. This pattern was compatible with an anti-leucine-rich glioma-inactivated 1 antibody encephalitis that was later confirmed.


Asunto(s)
Anticuerpos/inmunología , Fluorodesoxiglucosa F18 , Péptidos y Proteínas de Señalización Intracelular/inmunología , Encefalitis Límbica/diagnóstico por imagen , Tomografía Computarizada por Tomografía de Emisión de Positrones , Estudios de Seguimiento , Humanos , Encefalitis Límbica/inmunología , Masculino , Persona de Mediana Edad
20.
PLoS One ; 15(12): e0243729, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33315881

RESUMEN

This study aimed to directly analyze the potential relationship of anti-nuclear antibodies (ANA) before and after the administration of TNF-α inhibitors (TNFi) with the appearance of anti-drug antibodies (ADrA) in patients with rheumatoid arthritis (RA). A total of 121 cases, viz., 38, 53, and 30 cases treated with infliximab (IFX), adalimumab (ADA), and etanercept (ETN), respectively, were enrolled. The ANA titers were measured using indirect immunefluorescence assay (IF-ANA) and multiplex flow immunoassay (ANA Screen) before and serially during the therapy. The anti-IFX antibodies (HACA) and anti-ADA antibodies (AAA) were measured with a radioimmunoassay. ADrA turned positive in 14 (36.8%) among 38 patients treated with IFX, and 16 (30.2%) among 53 treated with ADA. All of them were positive for IF-ANA before TNFi administration, while ADrA never appeared in any of the 15 patients negative for IF-ANA (< 40). IF-ANA of high titers (≥ 320 and ≥ 640) before IFX treatment showed a significant association with the appearance of HACA 52 weeks after IFX (P = 0.040 and 0.017, respectively), whereas AAA appearance was not related to IF-ANA titers before treatment. Moreover, IF-ANA of high titers before IFX treatment was significantly associated with inefficacy and discontinuation of the treatment. The positivity of anti-SS-A antibodies before therapy might be a risk factor for ADrA appearance in patients treated with IFX or ADA. The percentage of patients whose IF-ANA titers increased was significantly higher with IFX than with ADA or ETN treatments (P = 0.026 and 0.022, respectively). High ANA titers and positive ANA Screen after IFX therapy showed a significant association with HACA appearance and possibly led to treatment failure. Among the three TNFi, only IFX showed a close relationship with IF-ANA and ADrA appearance, suggesting the interaction of immunogenicity with autoimmunity as well as the advantage of ANA measurement before TNFi therapy.


Asunto(s)
Adalimumab/inmunología , Anticuerpos/inmunología , Antirreumáticos/inmunología , Artritis Reumatoide/tratamiento farmacológico , Etanercept/inmunología , Infliximab/inmunología , Adalimumab/uso terapéutico , Adulto , Anciano , Anticuerpos Antinucleares/inmunología , Antirreumáticos/uso terapéutico , Artritis Reumatoide/inmunología , Etanercept/uso terapéutico , Femenino , Humanos , Infliximab/uso terapéutico , Masculino , Persona de Mediana Edad , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores
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