Immunogenomic engineering of a plug-and-(dis)play hybridoma platform
| LEADER | caa a22 4500 | ||
|---|---|---|---|
| 001 | 528791931 | ||
| 005 | 20190713071704.0 | ||
| 007 | cr unu---uuuuu | ||
| 008 | 180924s2016 xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.3929/ethz-b-000119720 |2 doi |
| 024 | 7 | 0 | |a 10.1038/ncomms12535 |2 doi |
| 035 | |a (ETHRESEARCH)oai:www.research-collecti.ethz.ch:20.500.11850/288673 | ||
| 035 | |a (ETHRESEARCH)oai:www.research-collecti.ethz.ch:20.500.11850/119720 | ||
| 245 | 0 | 0 | |a Immunogenomic engineering of a plug-and-(dis)play hybridoma platform |h [Elektronische Daten] |c [Mark Pogson, Cristina Parola, William J. Kelton, Paul Heuberger, Sai T. Reddy] |
| 246 | 0 | |a Nat Commun | |
| 506 | |a Open access |2 ethresearch | ||
| 520 | 3 | |a Hybridomas, fusions of primary mouse B cells and myelomas, are stable, rapidly-proliferating cell lines widely utilized for antibody screening and production. Antibody specificity of a hybridoma clone is determined by the immunoglobulin sequence of the primary B cell. Here we report a platform for rapid reprogramming of hybridoma antibody specificity by immunogenomic engineering. Here we use CRISPR-Cas9 to generate double-stranded breaks in immunoglobulin loci, enabling deletion of the native variable light chain and replacement of the endogenous variable heavy chain with a fluorescent reporter protein (mRuby). New antibody genes are introduced by Cas9-targeting of mRuby for replacement with a donor construct encoding a light chain and a variable heavy chain, resulting in full-length antibody expression. Since hybridomas surface express and secrete antibodies, reprogrammed cells are isolated using flow cytometry and cell culture supernatant is used for antibody production. Plug-and-(dis)play hybridomas can be reprogrammed with only a single transfection and screening step. | |
| 540 | |a Creative Commons Attribution 4.0 International |u http://creativecommons.org/licenses/by/4.0 |2 ethresearch | ||
| 700 | 1 | |a Pogson |D Mark |e joint author | |
| 700 | 1 | |a Parola |D Cristina |e joint author | |
| 700 | 1 | |a Kelton |D William J. |e joint author | |
| 700 | 1 | |a Heuberger |D Paul |e joint author | |
| 700 | 1 | |a Reddy |D Sai T. |e joint author | |
| 773 | 0 | |t Nature Communications |d London : Nature Publishing Group |g 7, p. 12535 |x 2041-1723 | |
| 856 | 4 | 0 | |u http://hdl.handle.net/20.500.11850/288673 |q text/html |z WWW-Backlink auf das Repository (Open access) |
| 908 | |D 1 |a Journal Article |2 ethresearch | ||
| 950 | |B ETHRESEARCH |P 856 |E 40 |u http://hdl.handle.net/20.500.11850/288673 |q text/html |z WWW-Backlink auf das Repository (Open access) | ||
| 950 | |B ETHRESEARCH |P 773 |E 0- |t Nature Communications |d London : Nature Publishing Group |g 7, p. 12535 |x 2041-1723 | ||
| 950 | |B ETHRESEARCH |P 700 |E 1- |a Pogson |D Mark |e joint author | ||
| 950 | |B ETHRESEARCH |P 700 |E 1- |a Parola |D Cristina |e joint author | ||
| 950 | |B ETHRESEARCH |P 700 |E 1- |a Kelton |D William J. |e joint author | ||
| 950 | |B ETHRESEARCH |P 700 |E 1- |a Heuberger |D Paul |e joint author | ||
| 950 | |B ETHRESEARCH |P 700 |E 1- |a Reddy |D Sai T. |e joint author | ||
| 950 | |B ETHRESEARCH |P 856 |E 40 |u http://hdl.handle.net/20.500.11850/119720 |q text/html |z WWW-Backlink auf das Repository (Open access) | ||
| 950 | |B ETHRESEARCH |P 773 |E 0- |t Nature Communications |d London : Nature Publishing Group |g 7, p. 12535 | ||
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 949 | |B ETHRESEARCH |F ETHRESEARCH |b ETHRESEARCH |j Journal Article |c Open access | ||
| 949 | |B ETHRESEARCH |F ETHRESEARCH |b ETHRESEARCH |j Journal Article |c Open access | ||