By-passing in vitro screening--next generation sequencing technologies applied to antibody display and in silico candidate selection.
In recent years, unprecedented DNA sequencing capacity provided by next generation sequencing (NGS) has revolutionized genomic research. Combining the Illumina sequencing platform and a scFv library designed to confine diversity to both CDR3, >1.9 × 10(7) sequences have been generated. This approach allowed for in depth analysis of the library's diversity, provided sequence information on virtually all scFv during selection for binding to two targets and a global view of these enrichment processes. Using the most frequent heavy chain CDR3 sequences, primers were designed to rescue scFv from the third selection round. Identification, based on sequence frequency, retrieved the most potent scFv and valuable candidates that were missed using classical in vitro screening. Thus, by combining NGS with display technologies, laborious and time consuming upfront screening can be by-passed or complemented and valuable insights into the selection process can be obtained to improve library design and understanding of antibody repertoires.
Ravn
U
U
NovImmune SA, Ch des Aulx 14 and Fasteris SA, 1228 Plan-les-Ouates, Switzerland.
Gueneau
F
F
Baerlocher
L
L
Osteras
M
M
Desmurs
M
M
Malinge
P
P
Magistrelli
G
G
Farinelli
L
L
Kosco-Vilbois
M H
MH
Fischer
N
N
D002460:Cell Line; D022801:Complementarity Determining Regions; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D006801:Humans; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D011786:Quality Control; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000235:genetics*
D022801:Complementarity Determining Regions; D019151:Peptide Library; D057127:Single-Chain Antibodies