title: Recombinant renewable polyclonal antibodies.
abstract: Only 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.
authors.affiliation: a New Mexico Consortium ; Los Alamos , NM USA.
authors.forename: Fortunato
authors.initials: F
authors.lastname: Ferrara
authors.forename: Sara
authors.initials: S
authors.lastname: D'Angelo
authors.forename: Tiziano
authors.initials: T
authors.lastname: Gaiotto
authors.forename: Leslie
authors.initials: L
authors.lastname: Naranjo
authors.forename: Hongzhao
authors.initials: H
authors.lastname: Tian
authors.forename: Susanne
authors.initials: S
authors.lastname: Gräslund
authors.forename: Elena
authors.initials: E
authors.lastname: Dobrovetsky
authors.forename: Peter
authors.initials: P
authors.lastname: Hraber
authors.forename: Fridtjof
authors.initials: F
authors.lastname: Lund-Johansen
authors.forename: Silvia
authors.initials: S
authors.lastname: Saragozza
authors.forename: Daniele
authors.initials: D
authors.lastname: Sblattero
authors.forename: Csaba
authors.initials: C
authors.lastname: Kiss
authors.forename: Andrew R M
authors.initials: AR
authors.lastname: Bradbury
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000235:genetics; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D000906:Antibodies; D000939:Epitopes; D011994:Recombinant Proteins
keywords: 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 poly; antibody validation; phage display; polyclonal recombinant antibodies; yeast display