RESUMO
Over the past few years, the SNAP-tag technology has become a methodology with great potential in a variety of applications, e.g. the (specific) visualization of individual proteins and studies of protein interaction in living cells. Furthermore, the tag can be used for immunopurification and detection of recombinant proteins or site-specific coupling of recombinant proteins to surfaces. Next to the in vitro applications, it also enables detection of tagged proteins in vivo. This review gives an overview of the SNAP-tag technology in different fields of research and its potential for future developments.
Assuntos
Imagem Molecular/métodos , Proteínas/química , Proteínas/metabolismo , Animais , Regulação da Expressão Gênica/fisiologia , Ligação Proteica , Proteínas/genética , Proteínas RecombinantesRESUMO
Biosensors are used for a variety of applications in medicine and biology. A critical step during the development of such devices is the coordination of biological and technical requirements. The design of the device, as well as of the sample chamber and its functionalized surface is of great importance. Depending on the surface, the method of coupling of the desired receptor has to be adapted to guarantee functionality and biological activity during the measuring process. By using the SNAP-tag technology, a site-specific coupling of molecules with unaltered activity to a variety of O(6)-benzylguanine functionalized surfaces is possible, making it a versatile tool for the setup of biomedical devices.
Assuntos
Técnicas Biossensoriais/instrumentação , Proteínas Imobilizadas/química , Imagem Molecular/métodos , Animais , Anticorpos , Proteínas Imobilizadas/metabolismo , Ligação ProteicaRESUMO
SNAP/CLIP-tag technology is a novel approach that allows tagged proteins to be covalently coupled to diverse labels, such as fluorochromes and particles, using a convenient and specific enzymatic reaction. A monoclonal antibody (mAb) that binds to the SNAP/CLIP-tag would be useful to determine labeling efficiency, and to achieve reproducible detection in a variety of experimental formats. We therefore generated the murine mAb M2D11 by standard immunization and hybridoma technology. M2D11 binds to both the SNAP- and the CLIP-tag in either the coupled or uncoupled configurations and can be detected in the context of ELISA, flow cytometry, immunohistochemistry and western blot. The new antibody increases the versatility of the SNAP-tag technology by enabling the detection of tagged proteins using conventional immunological methods and widely available secondary antibodies.