RESUMO
[In the present work virus particles of live mumps virus vaccine widely used for vaccination in Russia have been detected and visualized by atomic force microscopy. For quantitative estimation of the number of observed virus particles the special method has been proposed. The presence of protein component of the virus in vaccine was tested by ELISA and dot-blot analysis. Using quantitative real-time PCR assay the number of copies of viral RNA was estimated. The results of quantitative estimation obtained by real-time PCR corresponded with atomic force microscopy data.
Assuntos
Vacina contra Caxumba/química , Vacina contra Caxumba/genética , Vacina contra Caxumba/imunologia , Vírus da Caxumba/genética , Vírus da Caxumba/imunologia , Vírus da Caxumba/ultraestrutura , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Microscopia de Força Atômica/métodos , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
Specific interaction between human IgM and polyclonal antibodies immobilized on support was studied by atomic force microscopy. Human IgMs are responsible for a number of side effects arising during the xenotransplantation of mammalian organs to man. On the basis of atomic force microscopy, a quantitative analysis of complexes with IgM was performed. The data of the analysis agree well with the results of enzyme immunoassay. It was shown that the method of detection of immune complexes based on atomic force microscopy is able to detect specific antibodies/antigens in serum.
Assuntos
Complexo Antígeno-Anticorpo/análise , Imunoglobulina M/química , Silicatos de Alumínio/química , Animais , Anticorpos/química , Complexo Antígeno-Anticorpo/sangue , Bovinos , Humanos , Técnicas Imunoenzimáticas , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Ligantes , Microscopia de Força Atômica , Ligação ProteicaRESUMO
Complex formation between immunoglobulins and ligands immobilized on mica was studied by atomic force microscopy in two different systems. In the first system, 60-kDa ligands possessing only one site for antibody recognition were used. In the other system, a more complex interaction of human immunoglobulin with immobilized polyclonal antibodies was studied. In both systems, specific complexes with proper ligand appeared, and unspecific interaction was not detected. The method of revealing immunocomplexes by image atomic force microscopy can be used in the development of modern diagnostic systems.
Assuntos
Complexo Antígeno-Anticorpo/análise , Imunoglobulinas/química , Preparações de Plantas/química , Proteínas de Plantas/química , Ricina/química , Toxinas Biológicas/química , Silicatos de Alumínio/química , Anticorpos/química , Humanos , Imunoglobulina G/química , Imunoglobulinas/imunologia , Ligantes , Microscopia de Força Atômica , Peso Molecular , Preparações de Plantas/imunologia , Proteínas de Plantas/imunologia , Proteínas Inativadoras de Ribossomos Tipo 2 , Ricina/imunologia , Toxinas Biológicas/imunologiaRESUMO
Optimal conditions of protein scanning by atomic force microscopy were developed. Proteins of different molecular masses (950-11.5 kDa) and different three-dimensional organization were used to investigate the structural features of proteins. The most distinct images of proteins were obtained using a tip with the free amplitude in the range of 5-15 nm and with the set-point amplitude in the regime of repulsion from the sample. The method allowed one to clearly recognize the structural details of large molecules such as immunoglobulins IgM and IgG1 and Ricinus agglutinin. The revealing of the structural properties of proteins with molecular masses of 60 kDa and less was limited by the sharpness of probe tips used in the present study. It was shown that, by a quantitative analysis of the geometric parameters of molecules, it is possible to distinguish IgG1, Ricinus agglutinin, and ricin.
