Miniaturized parallelized sandwich immunoassays.

This chapter describes the development and use of bead-based miniaturized multiplexed sandwich immunoassays for focused protein profiling. Bead-based protein arrays or suspension microarrays allow simultaneous analysis of a variety of parameters within a single experiment. In suspension microarrays capture antibodies are coupled onto color-coded microspheres. The applications of suspension microarrays are described, which allow to analyze proteins present in different types of body fluids, such as serum or plasma, cerebrospinal, pleural and synovial fluids, as well as cell culture supernatants. The chapter is divided into the generation of suspension microarrays, sample preparation, processing of suspension microarrays, validation of analytical performance, and finally pattern generation using bioinformatics tools.

[1]  A I Saeed,et al.  TM4: a free, open-source system for microarray data management and analysis. , 2003, BioTechniques.

[2]  H. Horvitz,et al.  MicroRNA expression profiles classify human cancers , 2005, Nature.

[3]  U. Prabhakar,et al.  Simultaneous quantification of proinflammatory cytokines in human plasma using the LabMAP assay. , 2002, Journal of immunological methods.

[4]  Dieter Stoll,et al.  Miniaturised multiplexed immunoassays. , 2002, Current opinion in chemical biology.

[5]  W. Kuis,et al.  Improved multiplex immunoassay performance in human plasma and synovial fluid following removal of interfering heterophilic antibodies. , 2005, Journal of immunological methods.

[6]  Marta Sanchez-Carbayo,et al.  Antibody arrays: technical considerations and clinical applications in cancer. , 2006, Clinical chemistry.

[7]  S. Kingsmore.  Multiplexed protein measurement: technologies and applications of protein and antibody arrays , 2006, Nature Reviews Drug Discovery.

[8]  S. Dunbar,et al.  Quantitative, multiplexed detection of bacterial pathogens: DNA and protein applications of the Luminex LabMAP system. , 2003, Journal of microbiological methods.

[9]  A. Agadir,et al.  Cytometric bead array: a multiplexed assay platform with applications in various areas of biology. , 2004, Clinical immunology.

[10]  D. Vignali,et al.  Simultaneous quantitation of 15 cytokines using a multiplexed flow cytometric assay. , 1999, Journal of immunological methods.

[11]  J. W. Findlay,et al.  Validation of immunoassays for bioanalysis: a pharmaceutical industry perspective. , 2000, Journal of pharmaceutical and biomedical analysis.

[12]  R. Mage,et al.  A comparison of ELISA and flow microsphere-based assays for quantification of immunoglobulins. , 2002, Journal of immunological methods.

[13]  B. Natelson,et al.  Spinal Fluid Abnormalities in Patients with Chronic Fatigue Syndrome , 2005, Clinical Diagnostic Laboratory Immunology.

[14]  G. S. Johnson,et al.  An Information-Intensive Approach to the Molecular Pharmacology of Cancer , 1997, Science.