Screening flow cytometry

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Vetter, D., Tate, E.M., and Gallop, M.A. (1995) Strategies for the synthesis and screening of glycoconju-gates. 2. Covalent immobilization for flow cytometry. Bioconjugate Chem. 6, 319-322. 

Black CB, Duensing TD, Trinkle LS, Dunlay RT. Cell-based screening using high-throughput flow cytometry. Assay Drug Dev. Technol. 2010 Feb 9(l) 13-20. 

The second aspect of clinical practice that has led to a reassessment of the nature of flow cytometry is the occasional clinical requirement for rare-event analysis. Methods have been developed, particularly with the use of multiparameter gating, to lower background noise in order to provide increased sensitivity for detection of rare cells. In the clinic, this increased sensitivity translates, for example, into earlier diagnosis of relapse in leukemia, more sensitive detection of fetal-maternal hemorrhage, and better ability to screen leukocyte-reduced blood transfusion products for residual white blood cells. Outside the clinic, these methods for rare-event detection have begun to stretch the limits of research applications as well. 

The screening of protein libraries by flow cytometry offers the following advantages ... 

Unlike phage display and other screening technologies that rely on ligand binding, flow cytometry can be readily used to select clones either on the basis of binding or catalytic activity. Numerous enzymatic assays have already been adopted for use with flow cytometry and suitable commercial probes are available from vendors such as Molecular Probes. 

IV. Library Screening by Flow Cytometry A. Ligand Binding... 

Flow cytometry is well suited for the analysis of enzyme activity and kinetics at the single cell level (Watson and Dive, 1994). Flow cytometric assays for numerous enzymes including esterases, proteases, peroxidases, lipases, and oxidoreductases3 are available and are widely used in research and clinical practice. To date, flow cytometry has not been widely exploited as a screening tool for enzyme engineering purposes, but this is rapidly changing. 

MAbs produced by immunization with peptide may be screened first by ELISA to identify peptide specific antibodies. However, not all peptide specific antibodies will recognize the cell surface-expressed receptor. MAbs reactive with peptide must be subsequently screened by flow cytometry on receptor transfectants, cell lines, or leukocytes known to express the receptor in order to identify those which recognize native receptor. Alternatively, the fusion can be screened directly by flow cytometry to identify antibodies which recognize native receptor (see Subheading 3.3.2.),... 

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