title: Expression Cloning and Production of Human Heavy-Chain-Only Antibodies from Murine Transgenic Plasma Cells.
abstract: Several technologies have been developed to isolate human antibodies against different target antigens as a source of potential therapeutics, including hybridoma technology, phage and yeast display systems. For conventional antibodies, this involves either random pairing of VH and variable light (VL) domains in combinatorial display libraries or isolation of cognate pairs of VH and VL domains from human B cells or from transgenic mice carrying human immunoglobulin loci followed by single-cell sorting, single-cell RT-PCR, and bulk cloning of isolated natural VH-VL pairs. Heavy-chain-only antibodies (HCAbs) that naturally occur in camelids require only heavy immunoglobulin chain cloning. Here, we present an automatable novel, high-throughput technology for rapid direct cloning and production of fully human HCAbs from sorted population of transgenic mouse plasma cells carrying a human HCAb locus. Utility of the technique is demonstrated by isolation of diverse sets of sequence unique, soluble, high-affinity influenza A strain X-31 hemagglutinin-specific HCAbs.
authors.affiliation: Department of Cell Biology, Erasmus MC , Rotterdam , Netherlands.
authors.forename: Dubravka
authors.initials: D
authors.lastname: Drabek
authors.affiliation: Department of Cell Biology, Erasmus MC , Rotterdam , Netherlands.
authors.forename: Rick
authors.initials: R
authors.lastname: Janssens
authors.affiliation: Department of Cell Biology, Erasmus MC , Rotterdam , Netherlands.
authors.forename: Ernie
authors.initials: E
authors.lastname: de Boer
authors.affiliation: Harbour Antibodies BV , Rotterdam , Netherlands.
authors.forename: Rik
authors.initials: R
authors.lastname: Rademaker
authors.affiliation: WHO Influenza Centre, Frances Crick Institute , London , UK.
authors.forename: Johannes
authors.initials: J
authors.lastname: Kloess
authors.affiliation: WHO Influenza Centre, Frances Crick Institute , London , UK.
authors.forename: John
authors.initials: J
authors.lastname: Skehel
authors.affiliation: Department of Cell Biology, Erasmus MC, Rotterdam, Netherlands; Harbour Antibodies BV, Rotterdam, Netherlands.
authors.forename: Frank
authors.initials: F
authors.lastname: Grosveld
keywords: HCAb; HEK cell library; human VH; plasma cells; transgenic mice