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Presented by: Kristin Kuhn and Charlotte Yang

A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg , Annemarthe G van der Veen & Hidde L Ploegh. Presented by: Kristin Kuhn and Charlotte Yang. Monoclonal antibodies. Advantages

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Presented by: Kristin Kuhn and Charlotte Yang

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  1. A microengraving method for rapid selection of singlecells producing antigen-specific antibodiesJ Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg, Annemarthe G van derVeen& Hidde L Ploegh Presented by: Kristin Kuhn and Charlotte Yang

  2. Monoclonal antibodies • Advantages • Unlike polyclonal antibodies, they recognize only one epitope of an antigen • High specificity • Efficient binding • Less background noise • Disadvantages • Expensive to produce • Large time-scale necessary in order to isolate hybridomas • Current method: limiting serial dilution • Does not allow for high throughput analysis of secreted antibodies, or recovery of cells for clonal expansion

  3. Microengraving method • Created an array of microwells using photolithography and replica molding • Wells were 50/100 um in diameter and depth • Array was made of PDMS, a biocompatible material • Cells were deposited on surface • Excess was removed • # of cells dependent on concentration of cells, volume applied, time, size of wells, and size of slab • Achieved 1-3 cells in 50-75 % of the wells • Array is placed on top of a solid support (pre-treated).

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