生物化学技术

Breast cancer is one of the leading causes of death in women worldwide. During the last decade, great developments in our understanding of breast cancer at the molecular level have arisen from microarray data. Molecular profiling has supported the notion that breast cancer is not a simple disea ...

A large number of scoring functions for ranking peptide matches to observed MS/MS spectra have been discussed in the literature. In contrast to scoring functions, search strategies have received less attention, and an accurate description of search algorithms is limited. Proteomics is ...

High-throughput gene expression technologies based on DNA microarrays allow the examination of biological systems. However, the interpretation of the complex molecular descriptions generated by these approaches is still challenging. The development of new methodologies ...

The acylcarnitine profile is a diagnostic test for inherited disorders of fatty acid and branched-chain amino acid catabolism. Patients with this type of metabolic disorder accumulate disease-specific acylcarnitines that correlate with the acyl coenzyme A compounds in the affe ...

The diagnosis of inherited metabolic disorders of amino acid (AA) metabolism is based on the qualitative and/or the quantitative analysis of AAs, mainly in blood and urine. For years, the most widespread technique in use was ion-exchange chromatography followed by post-column derivati ...

Modern nuclear magnetic resonance (NMR) spectroscopy generates complex and information-rich metabolic profiles. These require robust, accurate, and often sophisticated statistical techniques to yield the maximum meaningful knowledge. In this chapter, we describe meth ...

High-resolution 1H magic angle spinning nuclear magnetic resonance (NMR), using a sample spinning rate of several kilohertz or more (i.e., high-resolution magic angle spinning (hr-MAS)), is a well-established method for metabolic profiling in intact tissues without the need for sample ...

Quantitative metabolic profiling originated as a 10-year project carried out between 1968 and 1978 in California. It was hypothesized and then demonstrated that quantitative analysis of a large number of metabolites – selected by analytical convenience and evaluated by computeri ...

Biofluids are by far the most commonly studied sample type in metabolic profiling studies, encompassing blood, urine, cerebrospinal fluid, cell culture media and many others. A number of these fluids can be obtained at a high sampling frequency with minimal invasion, permitting detailed c ...

The identification of drug metabolites in biofluids such as urine, plasma and bile is an important step in drug discovery and development. Proton nuclear magnetic resonance (1H-NMR) spectroscopy can provide detailed information regarding the structural transformation of a compo ...

Data processing forms a crucial step in metabolomics studies, impacting upon data output quality, analysis potential and subsequent biological interpretation. This chapter provides an overview of data processing and analysis of GC-MS- and LC-MS-based metabolomics data. Data pre ...

In the past decade, many new strategies for mass spectrometry (MS)-based analyses of lipids have been developed. Lipidomics is one of the most promising research fields to emerge as a result of these advances in MS. Currently, mass spectrometric analysis of lipids involves two complementary a ...

In this paper, analytical methodologies for the global profiling of lipids in serum and tissue samples are reported. The sample preparation is based on a modified Folch extraction, and the analysis is carried out with ultrahigh-performance liquid chromatography combined with mass spe ...

Metabolomics is a rapidly emerging field of functional genomics research whose aim is the comprehensive analysis of low molecular weight metabolites in a biological sample. Capillary electrophoresis–electrospray ionization-mass spectrometry (CE–ESI-MS) represents a p ...

Metabolomics involves qualitative and/or quantitative analysis of hundreds of metabolites in a complex sample. As most of the metabolites are polar in nature (for example, amino acids, nucleotides, carboxylic acids), their chromatographic separation and analysis present a diff ...

Metabolites produced by an organism can be quite extensive, and one analytical technique alone is not capable of their comprehensive detection and identification. The majority of environmental metabolomic studies have implemented proton nuclear magnetic resonance (1H-NMR) s ...

Metabolic fingerprinting, the main tool in metabolomics, is a non-targeted methodology where all detectable peaks (or signals), including those from unknown analytes, are considered to establish sample classification. After pattern comparison, those signals changing in resp ...

The identification of drug metabolites in biofluids such as urine, plasma, and bile, as well as in in vitro systems, is an important step in drug discovery and development. Mass spectrometry, particularly when combined with high-performance liquid chromatography (HPLC-MS), can enable de ...

Internal standards are extremely valuable for determining the pH and mol-wt ranges of a specific 2-D gel system and for constructing calibration curves for calculating the relative pI and M r of unknown proteins. Although external markers are often used for initially calibrating a 2-D gel, eve ...

The ATP-dependent degradation of ornithine decarboxylase is an exceptional case whereby a protein is targeted to the 26S proteasome independently of ubiquitin conjugation. Rather, prior association with the polyamine-induced regulatory protein, antizyme, confers suscep ...