(2015). Recombinant renewable polyclonal antibodies. mAbs: Vol. 7, No. 1, pp. 32-41. doi: 10.4161/19420862.2015.989047
AbstractOnly a small fraction of the antibodies in a traditional polyclonal antibody mixture recognize the target of interest, frequently resulting in undesirable polyreactivity. Here, we show that high-quality recombinant polyclonals, in which hundreds of different antibodies are all directed toward a target of interest, can be easily generated in vitro by combining phage and yeast display. We show that, unlike traditional polyclonals, which are limited resources, recombinant polyclonal antibodies can be amplified over one hundred million-fold without losing representation or functionality. Our protocol was tested on 9 different targets to demonstrate how the strategy allows the selective amplification of antibodies directed toward desirable target specific epitopes, such as those found in one protein but not a closely related one, and the elimination of antibodies recognizing common epitopes, without significant loss of diversity. These recombinant renewable polyclonal antibodies are usable in different assays, and can be generated in high throughput. This approach could potentially be used to develop highly specific recombinant renewable antibodies against all human gene products.
Via Krishan Maggon
mAbsVolume 7, Issue 1, 2015 Recombinant renewable polyclonal antibodies View full textDownload full textSupplementalFree accessDOI:10.4161/19420862.2015.989047Fortunato Ferraraa, Sara D’Angeloa, Tiziano Gaiottoaf, Leslie Naranjob, Hongzhao Tianb, Susanne Gräslundc, Elena Dobrovetskyc, Peter Hraberb, Fridtjof Lund-Johansend, Silvia Saragozzae, Daniele Sblatteroe, Csaba Kissb & Andrew RM Bradburyb*
pages 32-41
Publishing models and article dates explainedReceived: 11 Nov 2014Accepted: 13 Nov 2014Accepted author version posted online: 20 Dec 2014
Published online: 14 Jan 2015
Keywordspolyclonal recombinant antibodies, yeast display, phage display, antibody validation, CTBP, C-terminal binding protein; ELISA, enzyme linked immunosorbant assay; HCDR3, Heavy chain complementarity determining region 3; HPA, Human Protein Atlas; scFv, single chain Fv; PrESTs, Protein epitope signature tag; rrpAbs, recombinant renewable polyclonal antibodies; SDS–PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; TEV, tobacco etch virus