a. Overview. Monoclonal antibodies (mAbs) are antibodies of a single specificity derived from an immortalized B cell [251]. Human monoclonal antibodies can be made by applying traditional hybridoma technology to mice transgenic for the human immunoglobulin loci [252], by phage display [253], in which antibody fragments, such as single chain Fvs (scFv) or Fabs, are displayed on the surface of phage [254,255], or by yeast display [256- 258]. Naive phage antibody libraries, from which antibodies can be selected against most targets, are derived from natural unimmunized [255,259-262] or synthetic [263-266] rearranged (usually human) V genes, and are the commonest in vitro source used to derive antibody fragments. The use of phage display to select antibodies has two great advantages: first the ability to select antibodies with specific performance or recognition characteristics, and secondly, the fact that the gene for the antibody is cloned simultaneously with selection. This allows antibody fragment genes to be subjected to downstream engineering, including affinity maturation to affinities not attainable in natural immune systems [256,267-270], and reformatting to include effector functions for downstream applications [271-275], such as enzymatic activity (e.g. alkaline phosphatase) [272,275], immobilization tags for orientated coupling to surfaces [276,277], tandem affinity purification tags [278] for immunoprecipitation, dimerization domains [279], fluorescent labeling vectors [24,271,274] and recloning into the full length immunoglobulin format [38,280,281]. ScFvs have great advantages over conventional mAbs for STMC research, including their small size, useful for studies on the composition of signaling complexes at the membrane (Aims 1 and 2);their exquisite specificity, as needed to develop Abs that recognize specific phosphotyrosine sites on signaling proteins (Aim 1);and their monovalency, which is critical for the analysis of membrane dynamics (Aim 2). Technology for the generation of scFvs is limited to only a few labs woridwide. Thus the STMC Affinity Reagents Core, located in the Bradbury lab at LANL, is an enormous asset to the center.

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Chylek, Lily A; Akimov, Vyacheslav; Dengjel, Jörn et al. (2014) Phosphorylation site dynamics of early T-cell receptor signaling. PLoS One 9:e104240
Chylek, Lily A; Harris, Leonard A; Tung, Chang-Shung et al. (2014) Rule-based modeling: a computational approach for studying biomolecular site dynamics in cell signaling systems. Wiley Interdiscip Rev Syst Biol Med 6:13-36
Wu, Meiye; Piccini, Matthew E; Singh, Anup K (2014) MiRNA detection at single-cell resolution using microfluidic LNA flow-FISH. Methods Mol Biol 1211:245-60
Itano, Michelle S; Graus, Matthew S; Pehlke, Carolyn et al. (2014) Super-resolution imaging of C-type lectin spatial rearrangement within the dendritic cell plasma membrane at fungal microbe contact sites. Front Phys 2:
Lund, Troy C; Patrinostro, Xiaobai; Kramer, Ashley C et al. (2014) sdf1 Expression reveals a source of perivascular-derived mesenchymal stem cells in zebrafish. Stem Cells 32:2767-79
Liu, Sheng; Lidke, Keith A (2014) A multiemitter localization comparison of 3D superresolution imaging modalities. Chemphyschem 15:696-704
Steinkamp, Mara P; Low-Nam, Shalini T; Yang, Shujie et al. (2014) erbB3 is an active tyrosine kinase capable of homo- and heterointeractions. Mol Cell Biol 34:965-77
Westrate, Laura M; Drocco, Jeffrey A; Martin, Katie R et al. (2014) Mitochondrial morphological features are associated with fission and fusion events. PLoS One 9:e95265
Valley, Christopher C; Lidke, Keith A; Lidke, Diane S (2014) The spatiotemporal organization of ErbB receptors: insights from microscopy. Cold Spring Harb Perspect Biol 6:
Jerman, Stephanie; Ward, Heather H; Lee, Rebecca et al. (2014) OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts. PLoS One 9:e106330

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