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1.
Ann Lab Med ; 43(1): 86-91, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36045061

RESUMO

Antibodies against human CD36 are responsible for several immune-mediated disorders. The detection of anti-CD36 antibodies using the standard monoclonal antibody (mAb) immobilization of platelet antigens (MAIPA) assay is hampered by a high frequency of false-negative results, most likely due to competitive inhibition of the mAb used as the capture antibody. We generated a panel of mouse mAbs against CD36 and seven hybridomas (GZ-3, GZ-13, GZ-70, GZ-143, GZ-413, GZ-507, and GZ-608), which were selected for MAIPA assays, as they reacted with mouse and human CD36. Fourteen anti-CD36 sera were assayed; all of which showed a positive reaction in a PakPlus (Immucor GTI Diagnostics, Inc., Waukesha, WI, USA) ELISA-based screening (optical density: 0.257-2.292). When the reference anti-CD36 mAb FA6-152 was used in the MAIPA assay, only 6/14 (42.9%) sera displayed a positive reaction. In contrast, anti-CD36 antibodies were detected in 13/14 (92.9%) sera when GZ-70 and GZ-608 mAbs were used. This significant improvement resulted in the identification of anti-CD36 antibodies by an antigen capture assay. Since patient's platelets possibly carrying rare native antigens are used, this method will facilitate the identification of new platelet antibodies against CD36 that are involved in immune-mediated thrombocytopenia and other diseases, such as transfusion-related acute lung injury.


Assuntos
Antígenos de Plaquetas Humanas , Trombocitopenia , Animais , Anticorpos Monoclonais , Plaquetas , Antígenos CD36 , Humanos , Camundongos , Trombocitopenia/diagnóstico
2.
Food Chem ; 398: 133861, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35973297

RESUMO

Bisacodyl, sodium picosulfate and their metabolite bis-(p-hydroxyphenyl)-pyridyl-2-methane (BHPM) are clinically used to treat constipation. In this study, with the rational hapten design, a broad-specific and highly sensitive monoclonal antibody (mAb) was obtained with half of inhibitory concentrations of 0.16, 0.18 and 0.65 ng/mL for bisacodyl, sodium picosulfate and BHPM, respectively. Based on this mAb, a rapid and sensitive lateral flow immunochromatographic assay toward bisacodyl, sodium picosulfate and BHPM in slimming foods was developed. This method can qualitatively and quantitatively screen three stimulant laxatives with cut-off values of 3-6 ng/mL by naked eye and quantitative detection limits of 0.14-0.41 ng/mL by reader. The acceptable recoveries of spiked samples (78.00 %-120.12 %) and consistent results with liquid chromatography with tandem mass spectrometry (LC-MS/MS) in real sample detection confirmed the accuracy of the method. The established method provides a technique for multiplex, sensitive, fast, and on-site detection of three stimulant laxatives in slimming food.


Assuntos
Bisacodil , Laxantes , Anticorpos Monoclonais/química , Cromatografia de Afinidade/métodos , Cromatografia Líquida , Imunoensaio/métodos , Limite de Detecção , Espectrometria de Massas em Tandem
3.
Food Chem ; 400: 134012, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36055143

RESUMO

Exploring a novel strategy for strengthening the catalytic activity of enzyme facilitates the development of a sensitive enzyme-linked immunosorbent assay (ELISA). Herein, a chemical staining (CS) strategy was firstly discovered to possess the ability to directly improve the catalytic activity of horseradish peroxidase. Based on this discovery, coomassie brilliant blue was introduced into ELISA to establish a CS enhanced ELISA (CS-ELISA) to detect clenbuterol (CL) by simply staining monoclonal antibodies. Satisfactorily, the most important analytical parameters of CS-ELISA, including sensitivity (0.074 ng mL-1) and linear range (0.2-2 ng mL-1) were all improving 2-folds compared with conventional ELISA. Moreover, the CS-ELISA shows good applicability in the detection of CL in pork tenderloin samples. The proposed CS-ELISA shows various advantages, such as cost-effective, easily accessible, enhanced catalytic activity of enzyme, higher sensitivity, and broader linear range, providing a new insight into enhanced ELISA for food safety.


Assuntos
Clembuterol , Anticorpos Monoclonais , Clembuterol/análise , Ensaio de Imunoadsorção Enzimática , Peroxidase do Rábano Silvestre , Coloração e Rotulagem
4.
Food Chem ; 400: 134067, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36084594

RESUMO

To determine gentamicin residues in animal tissues, a monoclonal antibody (Mab) was produced and a sensitive indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) was developed. At first, gentamicin was conjugated with bovine serum albumin as immunogens which were used to immunize BALB/c mice. Then, an anti-gentamicin Mab was prepared by hybridoma technology. Finally, a sensitive icCLEIA was developed with an 50% inhibition concentration (IC50) of 0.067 ng/mL for gentamicin. The limit of detection of the icCLEIA was 0.002 ng/mL. The cross reactivity of the Mab with structural analogues were<0.01%. The recoveries of gentamicin ranged from 80 to 101% and coefficient of variation was <6.4% in pork and fish samples. Samples were detected by UPLC-MS/MS for evaluating reliability of the icCLEIA. The results suggested that the prepared anti-gentamicin Mab can be used for rapid and convenient immunoassays to detect gentamicin residues in animal tissues.


Assuntos
Anticorpos Monoclonais , Gentamicinas , Animais , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática/métodos , Imunoensaio/métodos , Técnicas Imunoenzimáticas , Luminescência , Camundongos , Camundongos Endogâmicos BALB C , Reprodutibilidade dos Testes , Soroalbumina Bovina , Espectrometria de Massas em Tandem
5.
MAbs ; 14(1): 2029675, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35133941

RESUMO

The functional interleukin 6 (IL-6) signaling complex is a hexameric structure composed of IL-6, IL-6Rα, and the signaling receptor gp130. There are three different modes of IL-6 signaling, classic signaling, trans-signaling, and trans-presentation, which are not functionally redundant and mediate pleiotropic effects on both physiological and pathophysiological states. Monoclonal antibodies against IL-6 or IL-6Rα have been successfully developed for clinical application. However, designing therapeutic interventions that block specific modes of IL-6 signaling in a pathologically relevant manner remains a great challenge. Here, we constructed a fusion protein Hyper-IL-6 (HyIL-6) composed of human IL-6 and IL-6Rα to develop specific blocking antibodies against the IL-6/IL-6Rα complex. We successfully screened the monoclonal antibody C14mab, which can bind to HyIL-6 with the binding constant 2.86 × 10-10 and significantly inhibit IL-6/IL-6Rα/gp130 complex formation. In vitro, C14mab effectively inhibited HyIL-6-stimulated signal transducer and activator of transcription 3 (STAT3) activation and related vascular endothelial growth factor (VEGF) induction. Moreover, C14mab efficaciously suppressed HyIL-6-induced acute phase response in vivo. Our data from hydrogen-deuterium exchange mass spectrometry demonstrate that C14mab mainly binds to site IIIa of IL-6 and blocks the final step in the interaction between gp130 and IL-6/IL-6Rα complex. Additionally, data from enzyme-linked immunosorbent assays and kinetics assays indicate that C14mab interacts simultaneously with IL-6 and IL-6Rα, while it does not interact with IL-6Rα alone. The unique features of C14mab may offer a novel alternative for IL-6 blockade and illuminate a better therapeutic intervention targeting IL-6.


Assuntos
Interleucina-6 , Receptores de Interleucina-6 , Anticorpos Monoclonais , Receptor gp130 de Citocina/química , Receptor gp130 de Citocina/metabolismo , Epitopos , Humanos , Interleucina-6/metabolismo , Receptores de Interleucina-6/química , Receptores de Interleucina-6/metabolismo , Fator A de Crescimento do Endotélio Vascular
6.
Mol Pharm ; 19(2): 508-519, 2022 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-34939811

RESUMO

Using light scattering (LS), small-angle X-ray scattering (SAXS), and coarse-grained Monte Carlo (MC) simulations, we studied the self-interactions of two monoclonal antibodies (mAbs), PPI03 and PPI13. With LS measurements, we obtained the osmotic second virial coefficient, B22, and the molecular weight, Mw, of the two mAbs, while with SAXS measurements, we studied the mAbs' self-interaction behavior in the high protein concentration regime up to 125 g/L. Through SAXS-derived coarse-grained representations of the mAbs, we performed MC simulations with either a one-protein or a two-protein model to predict B22. By comparing simulation and experimental results, we validated our models and obtained insights into the mAbs' self-interaction properties, highlighting the role of both ion binding and charged patches on the mAb surfaces. Our models provide useful information about mAbs' self-interaction properties and can assist the screening of conditions driving to colloidal stability.


Assuntos
Anticorpos Monoclonais , Anticorpos Monoclonais/química , Método de Monte Carlo , Espalhamento a Baixo Ângulo , Difração de Raios X , Raios X
7.
Gut Microbes ; 14(1): 2117503, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36100957

RESUMO

The origins of preexisting SARS-CoV-2 cross-reactive antibodies and their potential impacts on vaccine efficacy have not been fully clarified. In this study, we demonstrated that S2 was the prevailing target of the preexisting S protein cross-reactive antibodies in both healthy human and SPF mice. A dominant antibody epitope was identified on the connector domain of S2 (1147-SFKEELDKYFKNHT-1160, P144), which could be recognized by preexisting antibodies in both human and mouse. Through metagenomic sequencing and fecal bacteria transplant, we demonstrated that the generation of S2 cross-reactive antibodies was associated with commensal gut bacteria. Furthermore, six P144 reactive monoclonal antibodies were isolated from naïve SPF mice and were proven to cross-react with commensal gut bacteria collected from both human and mouse. A variety of cross-reactive microbial proteins were identified using LC-MS, of which E. coli derived HSP60 and HSP70 proteins were confirmed to be able to bind to one of the isolated monoclonal antibodies. Mice with high levels of preexisting S2 cross-reactive antibodies mounted higher S protein specific binding antibodies, especially against S2, after being immunized with a SARS-CoV-2 S DNA vaccine. Similarly, we found that levels of preexisting S2 and P144-specific antibodies correlated positively with RBD binding antibody titers after two doses of inactivated SARS-CoV-2 vaccination in human. Collectively, our study revealed an alternative origin of preexisting S2-targeted antibodies and disclosed a previously neglected aspect of the impact of gut microbiota on host anti-SARS-CoV-2 immunity.


Assuntos
COVID-19 , Microbioma Gastrointestinal , Vacinas Virais , Animais , Anticorpos Monoclonais , Anticorpos Antivirais , COVID-19/prevenção & controle , Vacinas contra COVID-19 , Escherichia coli , Humanos , Camundongos , SARS-CoV-2
8.
Medicine (Baltimore) ; 101(35): e30320, 2022 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-36107602

RESUMO

INTRODUCTION: Breast cancer (BC) is the most diagnosed cancer worldwide. Multiple myeloma (MM) is a hematologic malignancy characterized by the overproduction of monoclonal antibodies in the bone marrow. Systemic lupus erythematosus (SLE) is distinguished by the aberrant activity of the immune system with heterogeneous clinical manifestations. The coexistence of more than one major illness in a patient can present a diagnostic challenge for clinical physicians, especially when the comorbid diseases share a similar clinical presentation. Herein, we report an unusual case of secondary synchronous diagnosis of MM and SLE after BC treatment. PATIENT CONCERNS: A 69-year-old female patient with breast cancer experienced severe skin itching and rashes on the face, anterior chest wall, back, and trunk for two days before admission. She had high levels of immunoglobulin and anti-nuclear antibodies; low levels of complements 3 and 4; positive anti-cardiolipin-IgM, anti-beta 2 glycoprotein-1 (anti-ß2GP1) antibodies, and lupus anticoagulant results at serological testing. DIAGNOSIS: The postoperative pathology report showed ductal carcinoma in situ in the right breast. SLE was confirmed based on the 2019 European League Against Rheumatism/American College of Rheumatology (EULAR/ACR) criteria. IgG-κ type multiple myeloma was confirmed by bone marrow biopsy, and the patient was synchronously diagnosed with SLE and MM after BC treatment. INTERVENTIONS: Glucocorticoids and immunosuppressive agents, including intravenous hydrocortisone (5 g every 8 hours) and oral hydroxychloroquine (Plaquenil) (200 mg twice daily) were administered to treat SLE. One capsule of thalidomide 50 mg was administered orally every night at bedtime for MM. OUTCOMES: The patient died two days later, shortly after the administration of drugs, due to multiple organ failures secondary to pneumonia and respiratory failure. CONCLUSION: This is a case of MM and SLE after BC treatment. The present challenge was the early detection and accurate diagnosis of the secondary major illnesses, as the clinical manifestations were similar and non-specific between these two diseases. Awareness and prompt recognition of the common clinical symptoms of SLE and MM should be considered by clinical physicians to avoid delayed diagnoses and facilitate early treatment for a better prognosis.


Assuntos
Neoplasias da Mama , Lúpus Eritematoso Sistêmico , Mieloma Múltiplo , Idoso , Anticorpos Monoclonais/uso terapêutico , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/terapia , Feminino , Glicoproteínas/uso terapêutico , Humanos , Hidrocortisona/uso terapêutico , Hidroxicloroquina/uso terapêutico , Imunoglobulina G/uso terapêutico , Imunoglobulina M/uso terapêutico , Imunossupressores/uso terapêutico , Inibidor de Coagulação do Lúpus/uso terapêutico , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/diagnóstico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Mieloma Múltiplo/complicações , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/terapia , Talidomida/uso terapêutico
9.
Clin J Oncol Nurs ; 26(5): 479-482, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36108215

RESUMO

Tixagevimab-cilgavimab is the only nonvaccine drug currently authorized in the United States for the prevention of COVID-19 infection in individuals who are moderately to severely immunocompromised or unable to receive COVI.


Assuntos
COVID-19 , Infecções por HIV , Profilaxia Pré-Exposição , Anticorpos Monoclonais , COVID-19/prevenção & controle , Infecções por HIV/tratamento farmacológico , Humanos , Estados Unidos
10.
Cell Host Microbe ; 30(9): 1194-1195, 2022 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-36108609

RESUMO

Utilizing monoclonal antibodies to prevent and treat infectious diseases has been accelerated by the COVID-19 pandemic. In this issue of Cell Host & Microbe, Zheng et al. show how a three-monoclonal-antibody cocktail, that defies conventions of "rational design" for a therapeutic agent, functions cooperatively to disrupt coxsackievirus virions.


Assuntos
COVID-19 , Enterovirus , Anticorpos Monoclonais/uso terapêutico , Humanos , Pandemias , Vírion
11.
Sci Rep ; 12(1): 15496, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36109569

RESUMO

Since late 2019, the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the resultant spread of COVID-19 have given rise to a worldwide health crisis that is posing great challenges to public health and clinical treatment, in addition to serving as a formidable threat to the global economy. To obtain an effective tool to prevent and diagnose viral infections, we attempted to obtain human antibody fragments that can effectively neutralize viral infection and be utilized for rapid virus detection. To this end, several human monoclonal antibodies were isolated by bio-panning a phage-displayed human antibody library, Tomlinson I. The selected clones were demonstrated to bind to the S1 domain of the spike glycoprotein of SARS-CoV-2. Moreover, clone A7 in Fab and IgG formats were found to effectively neutralize the binding of S protein to angiotensin-converting enzyme 2 in the low nM range. In addition, this clone was successfully converted to quench-based fluorescent immunosensors (Quenchbodies) that allowed antigen detection within a few minutes, with the help of a handy fluorometer.


Assuntos
Bacteriófagos , Técnicas Biossensoriais , COVID-19 , Enzima de Conversão de Angiotensina 2 , Anticorpos Monoclonais , Anticorpos Neutralizantes , Anticorpos Antivirais , Bacteriófagos/metabolismo , COVID-19/diagnóstico , Humanos , Imunoensaio , Fragmentos de Imunoglobulinas , Imunoglobulina G , Glicoproteínas de Membrana/metabolismo , Biblioteca de Peptídeos , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Proteínas do Envelope Viral/metabolismo
12.
Sci Rep ; 12(1): 15517, 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36109550

RESUMO

Coronavirus disease 2019 (COVID-19) continues to significantly impact the global population, thus countermeasure platforms that enable rapid development of therapeutics against variants of SARS-CoV-2 are essential. We report use of a phage display human antibody library approach to rapidly identify neutralizing antibodies (nAbs) against SARS-CoV-2. We demonstrate the binding and neutralization capability of two nAbs, STI-2020 and STI-5041, against the SARS-CoV-2 WA-1 strain as well as the Alpha and Beta variants. STI-2020 and STI-5041 were protective when administered intravenously or intranasally in the golden (Syrian) hamster model of COVID-19 challenged with the WA-1 strain or Beta variant. The ability to administer nAbs intravenously and intranasally may have important therapeutic implications and Phase 1 healthy subjects clinical trials are ongoing.


Assuntos
COVID-19 , Animais , Anticorpos Monoclonais , Anticorpos Neutralizantes/uso terapêutico , Anticorpos Antivirais/uso terapêutico , Cricetinae , Humanos , Mesocricetus , Testes de Neutralização , SARS-CoV-2
13.
J Headache Pain ; 23(1): 123, 2022 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-36115947

RESUMO

BACKGROUND: In Italy, monoclonal antibodies targeting the CGRP pathway are subsidized for the preventive treatment of high frequency and chronic migraine (CM) in patients with a MIgraine Disability ASsessment (MIDAS) score ≥ 11. Eligibility to treatment continuation requires a ≥ 50% MIDAS score reduction at three months (T3). In this study, we evaluate whether a ≥ 50% MIDAS score reduction at T3 is a reliable predictor of response to one-year erenumab treatment. METHODS: In this prospective, open-label, real-world study, 77 CM patients were treated with erenumab 70-140 mg s.c. every 28 days for one year (T13). We collected the following variables: monthly migraine days (MMDs), monthly headache days (MHDs), days of acute medication intake, MIDAS, HIT-6, anxiety, depression, quality of life and allodynia. Response to erenumab was evaluated as: i) average reduction in MMDs during the 1-year treatment period; and ii) percentage of patients with ≥ 50% reduction in MMDs during the last 4 weeks after the 13th injection (RespondersT13). RESULTS: Erenumab induced a sustained reduction in MMDs, MHDs and intake of acute medications across the 12-month treatment period, with 64.9% of patients qualifying as RespondersT13. At T3, 55.8% of patients reported a ≥ 50% reduction in MIDAS score (MIDASRes) and 55.4% of patients reported a ≥ 50% reduction in MMDs (MMDRes). MIDASRes and MMDRes patients showed a more pronounced reduction in MMDs during the 1-year treatment as compared to NON-MIDASRes (MIDASRes: T0: 23.5 ± 4.9 vs. T13: 7.7 ± 6.2; NON- MIDASRes: T0: 21.6 ± 5.4 vs. T13: 11.3 ± 8.8, p = 0.045) and NON-MMDRes (MMDRes: T0: 23.0 ± 4.5 vs. T13: 6.6 ± 4.8; NON-MMDRes: T0: 22.3 ± 6.0 vs. T13: 12.7 ± 9.2, p < 0.001) groups. The percentage of RespondersT13 did not differ between MIDASRes (74.4%) and NON-MIDASRes (52.9%) patients (p = 0.058), while the percentage of RespondersT13 was higher in the MMDRes group (83.3%) when compared to NON-MMDRes (42.9%) (p = 0.001). MMDRes predicted the long-term outcome according to a multivariate analysis (Exp(B) = 7.128; p = 0.001), while MIDASRes did not. Treatment discontinuation based on MIDASRes would have early excluded 36.0% of RespondersT13. Discontinuation based on "either MIDASRes or MMDRes" would have excluded a lower percentage (16%) of RespondersT13. CONCLUSION: MIDASRes only partly reflects the 12-month outcome of erenumab treatment in CM, as it excludes more than one third of responders. A criterion based on the alternative consideration of ≥ 50% reduction in MIDAS score or MMDs in the first three months of treatment represents a more precise and inclusive option. TRIAL REGISTRATION: The trial was retrospectively registered at www. CLINICALTRIALS: gov (NCT05442008). CGRP: Calcitonin Gene Related Peptide. MIDAS: MIgraine Disability Assessment. MMDs: monthly migraine days. MIDASRes: Patients with a MIDAS score reduction of at least 50% at T3. MMDRes: Patients with a MMDs reduction of at least 50% at T3. ResponderT13: Patients with a MMDs reduction from baseline of at least 50% in the last 4 weeks of observation period (after 13 erenumab administrations). T0: First erenumab administration. T3, T6, T9, T12: Follow-up visits at three, six, nine, and twelve months after first erenumab administration. T13: Last visit of the protocol.


Assuntos
Peptídeo Relacionado com Gene de Calcitonina , Transtornos de Enxaqueca , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Peptídeo Relacionado com Gene de Calcitonina/uso terapêutico , Avaliação da Deficiência , Humanos , Transtornos de Enxaqueca/prevenção & controle , Estudos Prospectivos , Qualidade de Vida
14.
Biologicals ; 79: 10-18, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36085129

RESUMO

The ability of antibodies to distinctly identify the antigens is an important feature exploited by the scientific community for the treatment of various diseases. The therapeutic action of monoclonal antibodies (mAbs) is mediated along with the cells of the immune system, such as natural killer cells, T cells and macrophages. The two major mechanisms that govern the therapeutic efficacy of mAbs are the antibody dependent cell mediated cytotoxicity (ADCC) and the complement dependent cytotoxicity (CDC). Consequently, much of the research dedicated to improving their action is focussed on enhancing either of these mechanisms. This manuscript focuses on the strategies to enhance ADCC, for providing more efficacious mAb therapeutics. These approaches essentially bring about changes in the elements of ADCC mechanism, such as the effector cell or the antibody itself and thus favour an enhanced therapeutic response. Several technologies of ADCC enhancement have been developed, based on the success of various strategies advanced by the researchers. These technologies show success with a few antibody therapeutics while they do not work with others. This review presents a detailed overview on these strategies and presents perspectives regarding the same.


Assuntos
Citotoxicidade Celular Dependente de Anticorpos , Antineoplásicos Imunológicos , Anticorpos Monoclonais/uso terapêutico , Células Matadoras Naturais
15.
Biologicals ; 79: 19-26, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36096853

RESUMO

Canine morbillivirus is a highly contagious multi-host pathogen with high morbidity and mortality. Timely diagnosis is of utmost importance to effectively control such a dreadful disease. Monoclonal antibodies (mAbs) serve as a high throughput diagnostics and applied tools for research and development (R&D). In the present study, a total of six mouse monoclonal antibodies were developed. All the mAbs generated belonged to IgG class. Of the six mAbs, two of them, namely CD-2F8 and CD-3D8 were directed against the nucleocapsid protein of CDV as determined in western blotting. The reactivity of all the mAbs was checked in indirect-ELISA and cell-ELISA using different morbilliviruses. The mAbs could broadly be categorized as; CDV specific (CD-3D8 and CD-2F8), cross-reactive to PPR virus (CD-AB3 and CD-4D6) and cross-reactive to both PPR virus and measles virus (CD-5D10 and CD-6E5). The characterized mAbs were used for antigenic profiling of CDV, PPR virus and measles virus. Based on the reactivity pattern; a close antigenic relationship was found among CDV and PPR virus as compared to measles virus. A pair of CDV specific mAbs namely CD-2F8 and CD-3D8 were identified which did not cross-react with measles and PPR viruses and thus could be used for diagnostic applications.


Assuntos
Vírus da Cinomose Canina , Vírus da Peste dos Pequenos Ruminantes , Animais , Anticorpos Monoclonais , Cães , Imunoglobulina G , Vírus do Sarampo , Camundongos , Proteínas do Nucleocapsídeo
16.
Arch Osteoporos ; 17(1): 125, 2022 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-36114901

RESUMO

Denosumab leads to improvements in BMD levels and is a well-tolerated agent according to results of randomized controlled studies but results in real-life setting are important to evaluate drug adherence and real-life efficiency. In this study, we present the results of 305 patients that were treated with denosumab in our clinic. INTRODUCTION: The long-term efficacy of anti-osteoclastic drugs in treatment of osteoporosis is well known. Denosumab, a novel human monoclonal antibody, is an anti-osteoclastic agent that has been shown to lead to reductions in vertebral, nonvertebral, and hip fracture risk in randomized and observational studies. Real-life data of this agent is increasing. In this study, we presented our real-life data about the 2-year follow-up of patients under denosumab treatment. METHODS: Osteoporotic patients who were treated with at least one denosumab injection between 2014 and 2020 years were included. Clinical and demographic data, bone turnover markers, and radiological reports (bone mineral densitometry (BMD), vertebral x-ray) were obtained from patient files retrospectively. RESULTS: A total of 305 patients (f/m: 275/30, 68.1 ± 11.05 years) were included. The median injection number was 4 (1-10). Two hundred seventy-three patients (89.8%) were persistent on treatment at the 12th month; 175 patients (57.3%) were persistent at 24th month. Sixty-eight patients (22%) were not using denosumab anymore, 55 of the patients were not continuing by doctor desicion and 13 were not continuing due to patient-related causes. Median BMD levels significantly increased from 0.809 (0.2-1.601, IQR: 0.136) to 0.861 (0.517-1.607, IQR: 0.14) in L1-L4 and from 0.702 (0.349-0.997, IQR: 0.125) to 0.745 (0.508-1.008, IQR: 0.137) in femur area at the 24th month of treatment. An improvement of 8.04% in L1-L4 BMD and 4.5% in femur neck BMD levels at the 24th month of treatment was observed. There was a significant decrease in bone turnover markers at the 24th month of treatment. CONCLUSION: In our group of patients under denosumab treatment, 53% of persistence was found at 24 months and associated with improvement in BMD levels without any significant side effects except one case with urticarial reaction. Denosumab leads to improvements in BMD levels and is a well-tolerated agent in a real-life setting comparable to results of randomized controlled studies in patients with different comorbidities.


Assuntos
Conservadores da Densidade Óssea , Osteoporose Pós-Menopausa , Anticorpos Monoclonais , Densidade Óssea , Remodelação Óssea , Denosumab/uso terapêutico , Feminino , Seguimentos , Humanos , Osteoporose Pós-Menopausa/tratamento farmacológico , Estudos Retrospectivos
17.
PLoS One ; 17(9): e0273512, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36048906

RESUMO

Generating specific monoclonal antibodies (mAbs) that neutralize multiple antigen variants is challenging. Here, we present a strategy to generate mAbs that bind seven subtypes of botulinum neurotoxin serotype F (BoNT/F) that differ from each other in amino acid sequence by up to 36%. Previously, we identified 28H4, a mouse mAb with poor cross-reactivity to BoNT/F1, F3, F4, and F6 and with no detectable binding to BoNT/F2, F5, or F7. Using multicolor labeling of the different BoNT/F subtypes and fluorescence-activated cell sorting (FACS) of yeast displayed single-chain Fv (scFv) mutant libraries, 28H4 was evolved to a humanized mAb hu6F15.4 that bound each of seven BoNT/F subtypes with high affinity (KD 5.81 pM to 659.78 pM). In contrast, using single antigen FACS sorting, affinity was increased to the subtype used for sorting but with a decrease in affinity for other subtypes. None of the mAb variants showed any binding to other BoNT serotypes or to HEK293 or CHO cell lysates by flow cytometry, thus demonstrating stringent BoNT/F specificity. Multicolor FACS-mediated antibody library screening is thus proposed as a general method to generate multi-specific antibodies to protein subtypes such as toxins or species variants.


Assuntos
Anticorpos Monoclonais Humanizados , Toxinas Botulínicas , Citometria de Fluxo , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais Humanizados/química , Toxinas Botulínicas/imunologia , Reações Cruzadas , Citometria de Fluxo/métodos , Células HEK293 , Humanos , Camundongos , Anticorpos de Cadeia Única/química
18.
J Immunother Cancer ; 10(9)2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36104101

RESUMO

BACKGROUND: In this study, we describe the generation of a fully human monoclonal antibody (named '7NP2') targeting human fibroblast activation protein (FAP), an antigen expressed in the microenvironment of different types of solid neoplasms. METHODS: 7NP2 was isolated from a synthetic antibody phage display library and was improved by one round of mutagenesis-based affinity maturation. The tumor recognition properties of the antibody were validated by immunofluorescence procedures performed on cancer biopsies from human patients. A fusion protein consisting of the 7NP2 antibody linked to interleukin (IL)-12 was generated and the anticancer activity of the murine surrogate product (named mIL12-7NP2) was evaluated in mouse models. Furthermore, the safety of the fully human product (named IL12-7NP2) was evaluated in Cynomolgus monkeys. RESULTS: Biodistribution analysis in tumor-bearing mice confirmed the ability of the product to selectively localize to solid tumors while sparing healthy organs. Encouraged by these results, therapy studies were conducted in vivo, showing a potent antitumor activity in immunocompetent and immunodeficient mouse models of cancer, both as single agent and in combination with immune checkpoint inhibitors. The fully human product was tolerated when administered to non-human primates. CONCLUSIONS: The results obtained in this work provided a rationale for future clinical translation activities using IL12-7NP2.


Assuntos
Interleucina-12 , Neoplasias , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Humanos , Interleucina-12/metabolismo , Camundongos , Neoplasias/tratamento farmacológico , Distribuição Tecidual , Microambiente Tumoral
19.
Contrast Media Mol Imaging ; 2022: 1388517, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36105450

RESUMO

The objective of this study was to investigate the value of PET/CT imaging based on CNN image registration algorithm in evaluating the short-term efficacy of PD-1 monoclonal antibody immunotherapy for lymphoma. 36 patients with advanced lymphoma confirmed by histology or cytochemistry and treated with PD-1 monoclonal antibody admitted to hospital were included. In addition, 38 normal controls were from healthy volunteers. All patients were treated with PD-1 monoclonal antibody intravenous infusion, and the image data were processed by CT with intelligent segmentation algorithm. Medication method: nivolumab 3 mg/kg for 2 weeks; pembrolizumab 2 mg/kg for 3 weeks; and continuous medication until tumor progression or unacceptable adverse reactions. Efficacy was evaluated every 3 cycles. Imaging examinations were performed after 3 weeks of medication to evaluate the therapeutic effect. The concentrations of IL-2, IL-7, basic fibroblast growth factor (FGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), platelet-derived growth factor (PDGF-bb), and other factors in the normal control group were significantly higher than those in the advanced lymphoma patients, and the differences were statistically significant (P < 0.05). The PET/CT imaging automatic segmentation accuracy of the CNN image registration algorithm was greater than 81%, and 27 patients were treated for more than 3 cycles, including 1 case of partial remission (PR) (3.7%), 16 cases of stable disease (SD) (59.3%) after 3 cycles of treatment, and 10 cases of progressive disease (PD) (37%). After 6 cycles of treatment, 1 case was PR (8.3%), 7 cases were SD (58.4%), and 4 cases were PD (33.3%). Adverse reactions included fever, fatigue, gastrointestinal reactions, hypothyroidism, and interstitial pneumonia. PD-1 monoclonal antibody immunotherapy had a certain short-term effect on lymphoma.


Assuntos
Linfoma , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Algoritmos , Anticorpos Monoclonais/uso terapêutico , Humanos , Imunoterapia , Linfoma/diagnóstico por imagem , Linfoma/tratamento farmacológico , Redes Neurais de Computação , Tomografia por Emissão de Pósitrons , Receptor de Morte Celular Programada 1 , Tomografia Computadorizada por Raios X/métodos
20.
Int J Nanomedicine ; 17: 3989-4008, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36105615

RESUMO

Purpose: Ultrasound nanobubbles (NBs) can kill tumor cells, mediated by their effects of cavitation and acoustic perforation through ultrasound, while as novel drug carriers, biomaterial-modified NBs release drugs at a target region. In this work, the ultrasound NBs bridged by biotin-streptavidin were prepared simultaneously to be loaded with both programmed death ligand 1 monoclonal antibody (PD-L1 mAb) and doxorubicin (DOX), which are immune checkpoint inhibitors (ICIs) and chemotherapeutic agents, to synergize immunotherapy and chemotherapy combined with sonodynamic therapy (SDT). Methods: The PD-L1 mAb/DOX NBs, using bridging affinity biotin (BRAB) technology as a bridge, were prepared by thin-film hydration and mechanical oscillation for the targeted delivery of biotinylated PD-L1 mAb and DOX. Characterization and pharmacokinetic studies of PD-L1 mAb/DOX NBs were performed in vitro and in vivo. The antitumor effect of ultrasound-mediated PD-L1 mAb/DOX-NBs was studied in the subcutaneously transplanted tumor of the H22 hepatoma model, and the mechanism of synergistic tumor repression was investigated. Results: The data of in vitro targeting experiments, contrast-enhanced ultrasound imaging (CEUS), in vivo imaging of the small animals imaging system (IVIS), and frozen sections showed that PD-L1 mAb/DOX-NBs have well-targeted aggregation in the tumor. By observing tumor inhibition rate, tissue cell apoptosis, and apoptosis-related gene and protein expression, the PD-L1 mAb/DOX-NBs group showed the best immunotherapy effects, and its tumor volume and mass inhibition rates were about 69.64% and 75.97%, respectively (P < 0.01). Therefore, blocking the PD-1/PD-L1 pathway could improve immune cells' tumor-killing ability. Antitumor immune cytokines were further enhanced when combined with DOX-induced tumor cell apoptosis and immunogenic cell death (ICD). Conclusion: In summary, ultrasound-mediated PD-L1 mAb/DOX-NBs showed significant synergistic antitumor effects, providing a potential combined immunotherapy strategy for HCC.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Antígeno B7-H1 , Biotina , Carcinoma Hepatocelular/tratamento farmacológico , Linhagem Celular Tumoral , Doxorrubicina , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/tratamento farmacológico
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