| Title |
High Affinity Humanized Antibodies without Making Hybridomas; Immunization Paired with Mammalian Cell Display and In Vitro Somatic Hypermutation
|
|---|---|
| Published in |
PLOS ONE, November 2012
|
| DOI | 10.1371/journal.pone.0049458 |
| Pubmed ID | |
| Authors |
Audrey D. McConnell, Minjee Do, Tamlyn Y. Neben, Vladimir Spasojevic, Josh MacLaren, Andy P. Chen, Laurence Altobell, John L. Macomber, Ashley D. Berkebile, Robert A. Horlick, Peter M. Bowers, David J. King |
| Abstract |
A method has been developed for the rapid generation of high-affinity humanized antibodies from immunized animals without the need to make conventional hybridomas. Rearranged IgH D(J) regions were amplified from the spleen and lymph tissue of mice immunized with the human complement protein C5, fused with a limited repertoire of human germline heavy chain V-genes to form intact humanized heavy chains, and paired with a human light chain library. Completed heavy and light chains were assembled for mammalian cell surface display and transfected into HEK 293 cells co-expressing activation-induced cytidine deaminase (AID). Numerous clones were isolated by fluorescence-activated cell sorting, and affinity maturation, initiated by AID, resulted in the rapid evolution of high affinity, functional antibodies. This approach enables the efficient sampling of an immune repertoire and the direct selection and maturation of high-affinity, humanized IgGs. |
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Peru | 1 | 20% |
| Unknown | 4 | 80% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 4 | 80% |
| Science communicators (journalists, bloggers, editors) | 1 | 20% |
Mendeley readers
The data shown below were compiled from readership statistics for 93 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Brazil | 2 | 2% |
| Portugal | 1 | 1% |
| Sweden | 1 | 1% |
| Mexico | 1 | 1% |
| United States | 1 | 1% |
| Unknown | 87 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 35 | 38% |
| Student > Ph. D. Student | 26 | 28% |
| Student > Master | 9 | 10% |
| Student > Bachelor | 5 | 5% |
| Other | 4 | 4% |
| Other | 9 | 10% |
| Unknown | 5 | 5% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 41 | 44% |
| Biochemistry, Genetics and Molecular Biology | 27 | 29% |
| Chemistry | 4 | 4% |
| Immunology and Microbiology | 3 | 3% |
| Engineering | 3 | 3% |
| Other | 9 | 10% |
| Unknown | 6 | 6% |