High-throughput generation and engineering of recombinant human antibodies

Citation
B. Krebs et al., High-throughput generation and engineering of recombinant human antibodies, J IMMUNOL M, 254(1-2), 2001, pp. 67-84
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Immunology
Journal title
JOURNAL OF IMMUNOLOGICAL METHODS
ISSN journal
0022-1759 → ACNP
Volume
254
Issue
1-2
Year of publication
2001
Pages
67 - 84
Database
ISI
SICI code
0022-1759(20010801)254:1-2<67:HGAEOR>2.0.ZU;2-5
Abstract
The first version of the Human Combinatorial Antibody Library (HuCAL((R))) is a single-chain Fv-based phage display library (HuCAL((R))-scFv) with 2 X 10(9) members optimised for high-throughput generation and targeted engine ering of human antibodies. 61% of the library genes code for functional scF v as judged by sequencing. We show here that since HuCAL((R))-scFv antibodi es are expressed in high levels in Escherichia coli, automated panning and screening in miniaturised settings (96- and 384-well format) have now becom e feasible. Additionally, the unique modular design of HuCAL((R))-genes and -vectors allows the distinctly facilitated conversion of scFv into Fab, mi niantibody and immunoglobulin formats. and the fusion with a variety of eff ector functions and tags not only convenient for therapeutic applications b ut also for high-throughput purification and detection. Thus. the HuCAL((R) ) principle enables the rapid and high-throughput development of human anti bodies by optimisation strategies proven useful in classical low molecular weight drug development. We demonstrate in this report that HuCAL((R)) is a very convenient source of human antibodies for various applications. (C) 2 001 Elsevier Science B.V. All rights reserved.