RESUMO
Heparan sulfate (HS), a glycosaminoglycan present on the surface of cells, has been postulated to have important roles in driving both normal and pathological physiologies. The chemical structure and sulfation pattern (domain structure) of HS is believed to determine its biological function, to vary across tissue types, and to be modified in the context of disease. Characterization of HS requires isolation and purification of cell surface HS as a complex mixture. This process may introduce additional chemical modification of the native residues. In this study, we describe an approach towards thorough characterization of bovine kidney heparan sulfate (BKHS) that utilizes a variety of orthogonal analytical techniques (e.g. NMR, IP-RPHPLC, LC-MS). These techniques are applied to characterize this mixture at various levels including composition, fragment level, and overall chain properties. The combination of these techniques in many instances provides orthogonal views into the fine structure of HS, and in other instances provides overlapping / confirmatory information from different perspectives. Specifically, this approach enables quantitative determination of natural and modified saccharide residues in the HS chains, and identifies unusual structures. Analysis of partially digested HS chains allows for a better understanding of the domain structures within this mixture, and yields specific insights into the non-reducing end and reducing end structures of the chains. This approach outlines a useful framework that can be applied to elucidate HS structure and thereby provides means to advance understanding of its biological role and potential involvement in disease progression. In addition, the techniques described here can be applied to characterization of heparin from different sources.
Assuntos
Heparitina Sulfato/química , Animais , Bovinos , Cromatografia Líquida/métodos , Espectrometria de Massas/métodosRESUMO
Many vaccines, including those using recombinant antigen subunits, rely on adjuvant(s) to enhance the efficacy of the host immune responses. Among the few adjuvants clinically approved, QS-21, a saponin-based immunomodulatory molecule isolated from the tree bark of Quillaja saponaria (QS) is used in complex formulations in approved effective vaccines. High demand of the QS raw material as well as manufacturing scalability limitation has been barriers here. We report for the first-time successful plant cell culture production of QS-21 having structural, chemical, and biologic, properties similar to the bark extracted product. These data ensure QS-21 and related saponins are broadly available and accessible to drug developers.
RESUMO
Cyber security has become an important problem nowadays as almost everyone is often linked to the Internet for business and entertainment. Conventional cryptographers fail to address timely issues regarding cyber-attacks, such as cyber identity theft. In this work, we propose a novel idea, namely, a bandage-cover cryptographer (BCC), which is completely software-defined and protocol-free. Besides, this new cryptographic approach can enable camouflages to confuse data-mining robots, which are often encountered in the cyber world nowadays. Because all of the existing cryptographers aim to protect the entire data (document and file) altogether, they cannot have camouflagibility to mislead data-mining robots. Conversely, by our proposed novel BCC, one can select arbitrary contexts or parts of the data (related to individual identify and/or private confidential information) under protection. To evaluate such a first-ever cryptographer capable of misleading data-mining robots, we define two new metrics, namely: 1) vulnerability and 2) camouflage rates. The theoretical analyses of vulnerability rate and camouflage rate for our proposed new BCC are also presented in this article to demonstrate the corresponding effectiveness.