RESUMO
A novel recombinant single-chain fragment variable (scFv) antibody against Western equine encephalitis virus (WEE) was constructed and characterized. Using antibody phage display technology, a scFv was generated from the WEE specific hybridoma, 10B5 E7E2. The scFv was fused to a human heavy chain IgG1 constant region (CH1-CH3) and contained an intact 6 His tag and enterokinase recognition site (RS10B5huFc). The RS10B5huFc antibody was expressed in E. coli and purified by affinity chromatography as a 70-kDa protein. The RS10B5huFc antibody was functional in binding to WEE antigen in indirect enzyme-linked immunosorbent assays (ELISAs). Furthermore, the RS10B5huFc antibody was purified in proper conformation and formed multimers. The addition of the human heavy chain to the scFv replaced effector functions of the mouse antibody. The Fc domain was capable of binding to protein G and human complement. The above properties of the RS10B5huFc antibody make it an excellent candidate for immunodetection and immunotherapy studies.
Assuntos
Anticorpos Antivirais/química , Anticorpos Antivirais/genética , Vírus da Encefalite Equina do Oeste/imunologia , Fragmentos Fc das Imunoglobulinas/química , Fragmentos Fc das Imunoglobulinas/genética , Região Variável de Imunoglobulina/química , Região Variável de Imunoglobulina/genética , Proteínas Recombinantes/síntese química , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/isolamento & purificação , Anticorpos Antivirais/metabolismo , Antígenos Virais/imunologia , Antígenos Virais/metabolismo , Sítios de Ligação de Anticorpos , Clonagem Molecular , Humanos , Hibridomas , Fragmentos Fc das Imunoglobulinas/metabolismo , Região Variável de Imunoglobulina/metabolismo , Camundongos , Dados de Sequência Molecular , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismoRESUMO
This descriptive/exploratory study examined the concerns of families of patients with head and neck cancer. A convenience sample from a metropolitan hospital included four families. Data were collected through semistructured interviews, observations, and chart reviews over three time periods (before treatment, during treatment, and during rehabilitation) over a period of 5 months. Analysis of data showed five major types of concerns: cancer and its meaning, social relations, experience with hospitalization, treatment, and future placement. Findings showed that the types of concerns related to head and neck cancer are perceived as threatening and harmful. Concerns among patients were different from those of their family members across the different time periods. Our findings are useful in helping nurses understand the experience of head and neck cancer from the perspectives of the patient and their family members, and impact on their successful adjustment.
