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![Page 1: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/1.jpg)
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
![Page 2: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/2.jpg)
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
![Page 3: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/3.jpg)
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).
![Page 4: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/4.jpg)
![Page 5: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/5.jpg)
![Page 6: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/6.jpg)
![Page 7: A microengraving method for rapid selection of single cells producing antigen-specific antibodies J Christopher Love, Jehnna L Ronan, Gijsbert M Grotenbreg,](https://reader030.fdocuments.net/reader030/viewer/2022032723/56649f575503460f94c7c64e/html5/thumbnails/7.jpg)
Significance Can detect rare or slow-growing clones
due to early cell isolation Can perform screens with multiple
antigens Makes possible automation of monoclonal
antibody production