Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries
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Phage display of combinatorial antibody libraries is a very efficient method for selecting recombinant antibodies against a wide range of molecules. It has been applied very successfully for the generation of therapeutic antibodies for more than a decade. To increase robustness and reproducibility of the selection procedure, we developed a semi-automated selection method for the generation of recombinant antibodies from phage display libraries. In this procedure, the selection targets are specifically immobilised to magnetic particles which can then by automatically handled by a magnetic particle processor. At present up to 96 samples can be handled simultaneously. Applying the processor allows standardisation of panning parameters such as washing conditions, incubation times, or to perform parallel selections on same targets under different buffer conditions. Additionally, the whole protocol has been streamlined to carry out bead loading, phage selection, phage amplification between selection rounds and magnetic particle ELISA for confirmation of binding activity in microtiter plate formats. Until now, this method has been successfully applied to select antibody fragments against different types of target, such as peptides, recombinant or homologous proteins, or chemical compounds.
KeywordsMagnetic Bead Phage Display Antibody Fragment Phage Particle Antibody Phage
This work was supported by the German Federal Ministry for Education and Research (BMBF) through the National Genome Research Network (NGFN-II) project “Antibody Factory” (Grant No. 01GR0427) and the Max Planck Society. ZK acknowledges additional support from EU-FP6 CA “Proteome Binders” (RICA. 026008). TSL gratefully acknowledges financial support from the Ministry of Higher Education Malaysia and Institute for Research in Molecular Medicine, University Science Malaysia.
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