RESUMEN
Conformational changes of major histocompatibility complex (MHC) antigens have the potential to be recognized by T cells and may arise from polymorphic variation of the MHC molecule, the binding of modifying ligands, or both. Here, we investigated whether metal ions could affect allele-dependent structural variation of the two minimally distinct human leukocyte antigen (HLA)-B*27:05 and HLA-B*27:09 subtypes, which exhibit differential association with the rheumatic disease ankylosing spondylitis (AS). We employed NMR spectroscopy and X-ray crystallography coupled with ensemble refinement to study the AS-associated HLA-B*27:05 subtype and the AS-nonassociated HLA-B* 27:09 in complex with the self-peptide pVIPR (RRKWRRWHL). Both techniques revealed that pVIPR exhibits a higher degree of flexibility when complexed with HLA-B*27:05 than with HLA-B*27:09. Furthermore, we found that the binding of the metal ion Cu2+ or Ni2+, but not Mn2+, Zn2+, or Hg2+, affects the structure of a pVIPR-bound HLA-B*27 molecule in a subtype-dependent manner. In HLA-B*27:05, the metals triggered conformational reorientations of pVIPR, but no such structural changes were observed in the HLA-B*27:09 subtype, with or without bound metal ion. These observations provide the first demonstration that not only major histocompatibility complex class II, but also class I, molecules can undergo metal ion-induced conformational alterations. Our findings suggest that metals may have a role in triggering rheumatic diseases such as AS and also have implications for the molecular basis of metal-induced hypersensitivities and allergies.
Asunto(s)
Antígeno HLA-B27/química , Metales Pesados/química , Compuestos Organometálicos/química , Péptidos/química , Cristalografía por Rayos X , Antígeno HLA-B27/inmunología , Humanos , Metales Pesados/inmunología , Modelos Moleculares , Conformación Molecular , Compuestos Organometálicos/inmunología , Péptidos/inmunologíaAsunto(s)
Alérgenos/inmunología , Anafilaxia/diagnóstico , Medios de Contraste/administración & dosificación , Hipersensibilidad a las Drogas/diagnóstico , Cara/patología , Gadolinio/administración & dosificación , Meglumina/análogos & derivados , Compuestos Organometálicos/administración & dosificación , Adulto , Anafilaxia/etiología , Anafilaxia/prevención & control , Angioedema , Difenhidramina/uso terapéutico , Hipersensibilidad a las Drogas/complicaciones , Hipersensibilidad a las Drogas/tratamiento farmacológico , Femenino , Gadolinio/inmunología , Antagonistas de los Receptores Histamínicos H1/uso terapéutico , Humanos , Imagen por Resonancia Magnética , Meglumina/administración & dosificación , Meglumina/inmunología , Compuestos Organometálicos/inmunología , Pruebas Cutáneas , Síncope , UrticariaAsunto(s)
Medios de Contraste/efectos adversos , Reacciones Cruzadas/inmunología , Hipersensibilidad/inmunología , Compuestos Organometálicos/efectos adversos , Compuestos Organometálicos/inmunología , Gadolinio/efectos adversos , Gadolinio/inmunología , Humanos , Imagen por Resonancia Magnética/efectos adversos , Masculino , Persona de Mediana EdadAsunto(s)
Alérgenos/inmunología , Anafilaxia/diagnóstico , Aleteo Atrial/diagnóstico , Neoplasias Encefálicas/diagnóstico , Hipersensibilidad a las Drogas/diagnóstico , Gadolinio/inmunología , Glioma/diagnóstico , Angiografía por Resonancia Magnética , Compuestos Organometálicos/inmunología , Anciano , Algoritmos , Angioedema , Medios de Contraste , Epinefrina/administración & dosificación , Humanos , Hipotensión , Masculino , Persona de Mediana Edad , Pruebas Cutáneas , Triptasas/sangreRESUMEN
A series of zinc(II) dipicolylamine (ZnDPA)-based drug conjugates have been synthesized to probe the potential of phosphatidylserine (PS) as a new antigen for small molecule drug conjugate (SMDC) development. Using in vitro cytotoxicity and plasma stability studies, PS-binding assay, in vivo pharmacokinetic studies, and maximum tolerated dose profiles, we provided a roadmap and the key parameters required for the development of the ZnDPA based drug conjugate. In particular, conjugate 24 induced tumor regression in the COLO 205 xenograft model and exhibited a more potent antitumor effect with a 70% reduction of cytotoxic payload compared to that of the marketed irinotecan when dosed at the same regimen. In addition to the validation of PS as an effective pharmacodelivery target for SMDC, our work also provided the foundation that, if applicable, a variety of therapeutic agents could be conjugated in the same manner to treat other PS-associated diseases.
Asunto(s)
Antineoplásicos/farmacocinética , Inmunoconjugados/uso terapéutico , Terapia Molecular Dirigida/métodos , Compuestos Organometálicos/inmunología , Fosfatidilserinas/inmunología , Ácidos Picolínicos/inmunología , Animales , Antineoplásicos/síntesis química , Línea Celular Tumoral , Humanos , Inmunoconjugados/química , Inmunoconjugados/farmacocinética , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/farmacología , Fosfatidilserinas/metabolismo , Ácidos Picolínicos/síntesis química , Ácidos Picolínicos/farmacología , Relación Estructura-Actividad , Ensayos Antitumor por Modelo de XenoinjertoAsunto(s)
Alérgenos/inmunología , Hipersensibilidad a las Drogas/diagnóstico , Hipersensibilidad Tardía/diagnóstico , Linfocitos/inmunología , Compuestos Organometálicos/inmunología , Piel/patología , Biopsia , Medios de Contraste/química , Reacciones Cruzadas , Femenino , Gadolinio/química , Gadolinio/inmunología , Humanos , Imagen por Resonancia Magnética , Persona de Mediana Edad , Prednisolona/uso terapéuticoRESUMEN
Clinical status, hematological and biochemical parameters, and allergenic activity of organogermanium compounds used as adjuvants in complex with preparation from Orlov rubella virus vaccine strain and reference commercial anti-rubella vaccine based on Wistar RA 27/3 strain were studied on Macaca fascilcularis of both genders. Physiological parameters of monkeys immunized with the Russian and foreign rubella virus vaccine strains with and without adjuvants did not differ. The adjuvants were inessential for the safety of vaccines (absence of toxicity, reactogenic activity, or allergenic activity) in preclinical studies on lower primates.
Asunto(s)
Adyuvantes Inmunológicos/química , Anticuerpos Antivirales/sangre , Compuestos Organometálicos/inmunología , Vacuna contra la Rubéola/inmunología , Virus de la Rubéola/inmunología , Rubéola (Sarampión Alemán)/prevención & control , Adyuvantes Inmunológicos/administración & dosificación , Animales , Recuento de Células Sanguíneas , Evaluación Preclínica de Medicamentos , Femenino , Germanio/química , Germanio/inmunología , Inmunización , Inmunoglobulina E/sangre , Macaca fascicularis , Masculino , Compuestos Organometálicos/administración & dosificación , Compuestos Organometálicos/química , Ratas , Rubéola (Sarampión Alemán)/sangre , Rubéola (Sarampión Alemán)/inmunología , Rubéola (Sarampión Alemán)/virología , Vacuna contra la Rubéola/administración & dosificación , Vacuna contra la Rubéola/biosíntesis , Vacunas AtenuadasRESUMEN
AIM: Detailed characteristic of results of intranasal immunization of mice with one of two variants of vaccinating influenza virus, particularly in combination with a low molecular weight germanium-organic compound (LMW-GOC). An additional aim is evaluation of effect of LMW-GOC on the parameters of immune system in case of intranasal administration of the preparation without the addition of vaccinating virus. MATERIALS AND METHODS: The study was carried out in female CBA mice (18-20 g, 6 animals per group). Intranasal immunization was carried out by 2 different variants of B/Victoria influenza virus--once or twice with a 2 week interval. Cells for study were obtained from spleen and nasal- and bronchial-associated lymphoid tissue (NALT/ BALT) 24 hours and 7 days after intranasal administration of the preparations. The main method of the study--determination of the level of expression of various markers oflymphocytes in comparison with the level of the same markers in the cells of control group animals by using flow cytometry method. The mean parameters obtained were determined by using program package WinMDI 2.8. RESULTS: The main results were the increase of level of expression of various lymphocyte markers obtained from mice after intranasal administration of the vaccines and their combination with LMW-GOC or LMW-GOC only without the participation of the vaccines. A significant increase of the expression of TLR9 marker compared with other parameters was noted. Administration to mice of wild B/Victoria strain notably more frequently conditioned the decrease of expression of some parameters compared with administration of the cold adapted strain. Effect of LMW-GOC without the vaccine also conditioned the increase of levels of markers however a combination of the preparations with the vaccine was more effective. CONCLUSION: The increase of level of expression of a number of lymphocyte markers may serve as a sign of successful intranasal vaccination against influenza. LMW-GOC preparation increases immune stimulating effect of intranasally administered vaccines and in none of the cases weakens the stimulating result of effect of the vaccines, and in many cases increases it. LMW-GOC may be studied as a main or additional adjuvant for intranasal application of influenza vaccines.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Germanio , Virus de la Influenza B/inmunología , Vacunas contra la Influenza/inmunología , Linfocitos/efectos de los fármacos , Compuestos Organometálicos/administración & dosificación , Infecciones por Orthomyxoviridae/prevención & control , Adyuvantes Inmunológicos/síntesis química , Administración Intranasal , Animales , Anticuerpos Antivirales/sangre , Biomarcadores/metabolismo , Femenino , Expresión Génica , Humanos , Vacunas contra la Influenza/administración & dosificación , Linfocitos/inmunología , Ratones , Ratones Endogámicos CBA , Mucosa Nasal/inmunología , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/inmunología , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/virología , Receptor Toll-Like 9/biosíntesis , Receptor Toll-Like 9/inmunología , Vacunación , Vacunas AtenuadasRESUMEN
A homogeneous electrochemical immunoassay is based on the interaction of osmium-antigen conjugate with its antibody. The novelty presented herein is the direct conjugation of the osmium complex to a small antigen and the application of the quantitative analysis of the antigen and its antibody as the electrical signal for homogeneous immunoassay. The small antigen chosen is hippuric acid (HA), a major urinary metabolite in toluene-exposed humans. As a redox mediator, [Os(4,4'-dimethoxy-2,2'-bipyridine)2(4-aminomethylpyridine-HA)Cl](+/2+) (Os-HA antigen) has been synthesized and characterized on screen-printed carbon electrodes. The synthesized Os-HA antigen shows reversible redox peaks at E(½)=0.056 V versus Ag/AgCl. The homogeneous competitive immunoassay relies on the interaction between Os-HA antigen conjugate and free antigen to its antibody, which can generate electrical signals linearly proportional to the free antigen monitored by cyclic voltammetry and differential pulse voltammetry in the range of 10 µg mL(-1) to 5.12 mg mL(-1). The cutoff concentration of HA in urine samples is 2.0 mg mL(-1), so the method can be used to develop a HA immunosensor. Moreover, the proposed homogeneous electrochemical immunoassay method can be applied to detect low concentrations of small antigens found in the healthcare area.
Asunto(s)
Técnicas Electroquímicas , Hipuratos/orina , Inmunoensayo , Compuestos Organometálicos/química , Osmio/química , Animales , Reacciones Antígeno-Anticuerpo , Antígenos/inmunología , Antígenos/orina , Carbono/química , Electrodos , Hipuratos/inmunología , Humanos , Ratones , Ratones Endogámicos BALB C , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/inmunología , Osmio/inmunología , Propiedades de SuperficieRESUMEN
The organic germanium compound, Ge-132, has immune-modulating effects. We evaluated the symbiotic effects of Ge-132 with lactobacilli and oligosaccharide (LB/OS) on the immune responses of mice. The highest fecal IgA levels were observed in the mice receiving a low concentration of Ge-132 with LB/OS for 8 weeks. Our data suggest that LB/OS with a low concentration of Ge-132 stimulated the intestinal immunity.
Asunto(s)
Germanio/inmunología , Inmunidad/efectos de los fármacos , Inmunomodulación , Lactobacillus/inmunología , Oligosacáridos/inmunología , Compuestos Organometálicos/inmunología , Animales , Sinergismo Farmacológico , Germanio/farmacología , Inmunoglobulina A/análisis , Inductores de Interferón , Intestinos/inmunología , Ratones , Oligosacáridos/farmacología , Compuestos Organometálicos/farmacología , PropionatosRESUMEN
A Fischer carbene complex was grafted onto self-assembled monolayers (SAMs) on gold or glass by a copper-free "click" reaction. Pendant lysine residues of protein A obtained from Staphylococcus aureus rapidly reacted with the electrophilic metal complex on SAM effecting a covalent attachment of protein A with the surface. The protein A coated surface further led to bioaffinity immobilization of rabbit IgG in an oriented manner, a feature that also permits its purification from rabbit serum. Rabbit IgG could be removed from protein A coated surface by pH adjustment. The regenerated protein A surface was reused three times without loss of activity.
Asunto(s)
Inmunoglobulina G/inmunología , Membranas Artificiales , Metano/análogos & derivados , Compuestos Organometálicos/inmunología , Proteína Estafilocócica A/inmunología , Animales , Química Clic , Vidrio/química , Oro/química , Concentración de Iones de Hidrógeno , Inmunoglobulina G/sangre , Inmunoglobulina G/aislamiento & purificación , Metano/química , Estructura Molecular , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/química , Tamaño de la Partícula , Conejos , Proteína Estafilocócica A/química , Staphylococcus aureus/química , Estereoisomerismo , Propiedades de Superficie , Tungsteno/químicaRESUMEN
A detailed method for preparation of 2-mercaptobenzothiazole (MBT)-labeled immuno-Au aggregates as a novel Raman probe for SERS-based immunoassay has been proposed in this paper. The formation kinetics of gold aggregates induced by MBT and the impact of the amount of labeled reporter on the aggregation, as well as the fabrication of MBT-labeled immuno-Au aggregates were characterized comprehensively by UV-vis spectrophotometer. Meanwhile, a chain-like morphology of aggregates was monitored by TEM images. Experimental results show that this Raman tag can act as an efficient label and an enhanced Raman signal of SERS-based immunoassay can be obtained successfully with this novel probe. This work shows the method for the preparation of immune probe for SERS-based immunoassay using MBT as reporter and provides a reference for the preparation of immune probe with other Raman markers for multi-channel SERS-based immunoassay.
Asunto(s)
Benzotiazoles/química , Oro/química , Inmunoensayo/métodos , Espectrometría Raman/métodos , Oro/inmunología , Cinética , Microscopía Electrónica de Transmisión , Sondas Moleculares/química , Sondas Moleculares/inmunología , Sondas Moleculares/ultraestructura , Compuestos Organometálicos/química , Compuestos Organometálicos/inmunología , Espectrofotometría/métodosRESUMEN
Forty-four adult patients with cutaneous leishmaniasis (CL) were enrolled in a randomized, double-blind, controlled, dose-escalating clinical trial and were randomly assigned to receive three injections of either the LEISH-F1+MPL-SE vaccine (consisting of 5, 10, or 20 µg recombinant Leishmania polyprotein LEISH-F1 antigen+25 µg MPL-SE adjuvant) (n=27), adjuvant alone (n=8), or saline placebo (n=9). The study injections were given subcutaneously on Days 0, 28, and 56, and the patients were followed through Day 336 for safety, immunological, and clinical evolution endpoints. All patients received chemotherapy with meglumine antimoniate starting on Day 0. The vaccine was safe and well tolerated. Nearly all vaccine recipients and no adjuvant-alone or placebo recipients demonstrated an IgG antibody response to LEISH-F1 at Day 84. Also at Day 84, 80% of vaccine recipients were clinically cured, compared to 50% and 38% of adjuvant-alone and placebo recipients. The LEISH-F1+MPL-SE vaccine was safe and immunogenic in CL patients and appeared to shorten their time to cure when used in combination with meglumine antimoniate chemotherapy.
Asunto(s)
Leishmaniasis Cutánea/terapia , Meglumina/administración & dosificación , Compuestos Organometálicos/administración & dosificación , Vacunas Antiprotozoos/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Adolescente , Adulto , Anticuerpos Antiprotozoarios/sangre , Formación de Anticuerpos , Antígenos de Protozoos/inmunología , Método Doble Ciego , Quimioterapia Combinada , Femenino , Humanos , Inmunoglobulina G/sangre , Leishmaniasis Cutánea/inmunología , Masculino , Meglumina/inmunología , Antimoniato de Meglumina , Persona de Mediana Edad , Compuestos Organometálicos/inmunología , Poliproteínas/inmunología , Vacunas Antiprotozoos/efectos adversos , Proteínas Recombinantes/inmunología , Adulto JovenRESUMEN
Probe-capture systems based on proteins and synthetic ligands have become important for new analytical and imaging applications. We have used kinetic measurements of luminescence and measurements of binding by isothermal calorimetry to determine essential rate and equilibrium constants for a system that permanently captures modified DOTA chelates for positron imaging. We used that information along with previous results to quantitatively characterize the behavior of this system in vitro and in vivo. Under physiological conditions at 37 degrees C, the equilibrium dissociation constant for yttrium S-2-(4-aminobenzyl)-1,4,7,10-tetraazacyclododecanetetraacetate from antibody 2D12.5 is 2.0 (+/- 0.4) x 10(-9) M and the dissociation rate constant is 7.0 (+/- 0.7) x 10(-3) s(-1), leading to an inferred association rate constant of 3.5 x 10(6) M(-1) s(-1). Using these values to interpret data from earlier experiments leads to the rate constant 2.5 x 10(-2) s(-1) for covalent attachment of bound yttrium S-2-(4-acrylamidobenzyl)-1,4,7,10-tetraazacyclododecanetetraacetate to the G54C mutant of antibody 2D12.5. These values lead to a model for the detailed behavior of the latter system for tumor imaging in vivo that is consistent with experimental observations.
Asunto(s)
Anticuerpos/química , Anticuerpos/inmunología , Afinidad de Anticuerpos , Sondas Moleculares/química , Sondas Moleculares/inmunología , Calorimetría , Cinética , Luminiscencia , Mediciones Luminiscentes , Compuestos Organometálicos/química , Compuestos Organometálicos/inmunología , Tomografía de Emisión de Positrones , Termodinámica , Factores de TiempoRESUMEN
Dendritic cells play a pivotal role in host immune defense, such as elimination of foreign pathogen and inhibition of tumorigenesis. In this paper, we report that [Gd@C(82)(OH)(22)](n) could induce phenotypic maturation of dendritic cells by stimulating DC production of cytokines including IL-12p70, upregulating DC co-stimulatory (CD80, CD83, and CD86) and MHC (HLA-A,B,C and HLA-DR) molecules, and switching DCs from a CCL5-responsive to a CCL19-responsive phenotype. We found that [Gd@C(82)(OH)(22)](n) can induce dendritic cells to become functionally mature as illustrated by their capacity to activate allogeneic T cells. Mice immunized with ovalbumin in the presence of [Gd@C(82)(OH)(22)](n) exhibit enhanced ovalbumin-specific Th1-polarized immune response as evidenced by the predominantly increased production of IFNgamma, IL-1beta, and IL-2. The [Gd@C(82)(OH)(22)](n) nanoparticle is a potent activator of dendritic cells and Th1 immune responses. These new findings also provide a rational understanding of the potent anticancer activities of [Gd@C(82)(OH)(22)](n) nanoparticles reported previously.
Asunto(s)
Células Dendríticas/citología , Fulerenos/química , Gadolinio/química , Nanopartículas/química , Compuestos Organometálicos/inmunología , Compuestos Organometálicos/farmacología , Células TH1/inmunología , Animales , Antígenos/inmunología , Antineoplásicos/química , Antineoplásicos/inmunología , Antineoplásicos/farmacología , Diferenciación Celular/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Células Dendríticas/metabolismo , Humanos , Interleucina-6/biosíntesis , Ratones , Compuestos Organometálicos/químicaAsunto(s)
Anafilaxia/inducido químicamente , Medios de Contraste/efectos adversos , Compuestos Heterocíclicos/efectos adversos , Compuestos Organometálicos/efectos adversos , Anafilaxia/inmunología , Femenino , Gadolinio , Compuestos Heterocíclicos/inmunología , Humanos , Persona de Mediana Edad , Compuestos Organometálicos/inmunologíaRESUMEN
PURPOSE: Melanin has emerged as an attractive target for radioimmunotherapy (RIT) of melanoma, and a radiolabeled monoclonal antibody (mAb) 6D2 to melanin is currently in clinical evaluation. We investigated two approaches to improve the targeting of radiation to tumors using melanin-binding mAbs: (a) the use of an additional mAb to melanin could provide information on whether using antibodies to melanin can serve as a general approach to development of therapeutics for melanoma, and (b) as melanin targeting involves the antibody binding to extracellular melanin released from necrotic melanoma cells, we hypothesized that the administration of a chemotherapeutic agent followed by RIT would facilitate the delivery of radiation to the tumors due to the increased presence of free melanin. EXPERIMENTAL DESIGN: We evaluated the therapeutic efficacy of two melanin-binding IgM mAbs labeled with (188)Re (6D2 and 11B11). We compared the efficacy of RIT with (188)Re-6D2 to chemotherapy with dacarbazine (DTIC) and to combined chemotherapy and RIT in human metastatic melanoma-bearing nude mice. RESULTS: Therapeutic efficacy of (188)Re-labeled 6D2 and 11B11 was comparable despite differences in their affinity and binding site numbers. Comparison of chemotherapy with DTIC and RIT revealed that RIT was more effective in slowing tumor growth in mice. Administration of DTIC followed by RIT was more effective than either modality alone. CONCLUSIONS: These results provide encouragement for the development of RIT for melanoma with melanin-binding mAbs and suggest that combining chemotherapy and RIT may be a promising approach for the treatment of metastatic melanoma.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Antineoplásicos Alquilantes/uso terapéutico , Dacarbazina/uso terapéutico , Melaninas/inmunología , Melanoma/terapia , Radioinmunoterapia , Neoplasias Cutáneas/terapia , Animales , Anticuerpos Monoclonales/inmunología , Línea Celular Tumoral , Terapia Combinada , Femenino , Humanos , Melaninas/análisis , Melanoma/tratamiento farmacológico , Melanoma/radioterapia , Ratones , Ratones Desnudos , Metástasis de la Neoplasia , Compuestos Organometálicos/inmunología , Compuestos Organometálicos/uso terapéutico , Tomografía de Emisión de Positrones , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/radioterapiaRESUMEN
5'- N-Phenylacetyl sTn (sTnNPhAc), an unnatural derivative of sTn antigen expressed by many tumors, and its alpha-linked protein conjugates were prepared and investigated to explore glycoconjugate cancer vaccines. sTnNPhAcalpha-KLH elicited a robust T cell dependent immunity. The antiserum derived from sTnNPhAcalpha- or sTnNPhAcbeta-KLH-inoculated mice was similarly reactive to sTnNPhAcalpha and sTnNPhAcbeta but showed very little reactivity to sTn, NeuNPhAcalpha(2,3)GalNAc--a regioisomer of sTnNPhAc, isolated phenylacetyl group, and the linker employed to conjugate sTnNPhAc and carrier protein. It was concluded that the sTnNPhAc-elicited immunity was specific for the whole antigen rather than the phenylacetyl group or other partial structures of sTnNPhAc and that the reducing end configuration or linkage of sTnNPhAc did not affect its immunological identity. It was also concluded that a new linker designed to conjugate carbohydrates and proteins did not provoke any immune reaction and that the linker, as well as the associated new and convenient coupling strategy, can be safely used for the development of glycoconjugate vaccines.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores/metabolismo , Vacunas contra el Cáncer/metabolismo , Proteínas Portadoras/metabolismo , Glicoconjugados/metabolismo , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/inmunología , Animales , Antígenos de Carbohidratos Asociados a Tumores/química , Antígenos de Carbohidratos Asociados a Tumores/inmunología , Vacunas contra el Cáncer/inmunología , Glicoconjugados/inmunología , Hemocianinas/metabolismo , Ratones , Compuestos Organometálicos/química , Compuestos Organometálicos/metabolismo , Albúmina Sérica/metabolismo , EstereoisomerismoRESUMEN
An immunoliposome (80 nm in diameter) encapsulating a Ru complex with two aminobutyl moieties was prepared to detect the presence of hemagglutinin molecules, which play an important role in influenza virus infection. The highly sensitive detection was accomplished by electrochemiluminescence (ECL) from the Ru complex adsorbed onto Au electrodes after competitive immunoreactions. This method clarified that the adsorption of the Ru complex onto the electrode was an important factor in obtaining high sensitivity. Optimization of the analytical conditions enabled determination of the hemagglutinin molecules of the influenza virus in the concentration range of 3 x 10(-14) (6 x 10(-19) mol/50 microL sample) to 2 x 10(-12) g/mL. The sensitivity was far superior to that obtained by conventional ELISA as well as to that obtained by biosensors and reported thus far.
Asunto(s)
Técnicas Biosensibles/instrumentación , Hemaglutininas/análisis , Inmunoensayo/métodos , Liposomas/inmunología , Mediciones Luminiscentes/métodos , Compuestos Organometálicos/inmunología , Orthomyxoviridae/química , Compuestos de Rutenio/inmunología , Secuencia de Aminoácidos , Anticuerpos Monoclonales/inmunología , Calibración , Electroquímica , Electrodos , Oro/química , Hemaglutininas/inmunología , Datos de Secuencia Molecular , Compuestos Organometálicos/química , Orthomyxoviridae/inmunología , Orthomyxoviridae/aislamiento & purificación , Péptidos/química , Compuestos de Rutenio/química , Sensibilidad y EspecificidadRESUMEN
Recently, PET has been paid a great deal of attention as a non-invasive imaging method. In this review, the recent advances of PET using biomolecules, such as peptides, monoclonal antibodies, proteins, oligonucleotides, and glycoproteins will be described. So far, PET of biomolecules has been mainly used for diagnosis of cancers. The biomolecules have been conjugated with the DOTA ligand, labeled with radiometals as the beta+ emitter, and targeted to specific tumors, where they have enabled visualization of even small metastatic lesions, due to the high sensitivity of the PET scanners. Some of the biomolecules have been used not only for PET diagnosis, but also for radiotherapeutic treatments by simply changing the radiometals to beta(-) emitters. Collaborative work between chemists, biologists, and physicians will be important for the future of biomolecule-based targeting and diagnosis.