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A bioactivity-guided FFLP-based strategy for purification of a recombinant antibody Fc fragment. A strategy was developed to exploit the characteristics of an antigen-binding fragment of an antibody (a fragment of variable antibody light chain [V(L)]) to purify an IgG1. Using an Escherichia coli expression system, a V(L) encoding a recombinant Fab fragment (rFab) was first expressed and then solubilized in mild conditions that did not interfere with the function of the rFab. The rFab was then selectively purified by affinity chromatography using a phage display library in which the gene for the variable heavy (V(H)) region of the same antibody was displayed on the surface of the phage. Affinity selection of rFab-displaying phage was optimized using a biopanning step in which the V(H) was immobilized as a reference molecule. This purification strategy was applied to the isolation of a functional rFab from which the IgG1 was expressed and characterized. The purity of the rFab was verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoaffinity assays. The ability of the rFab to bind antigen was investigated by surface plasmon resonance (SPR). Finally, the biological activity of the rFab was confirmed by enzyme-linked immunosorbent assays and SPR. In conclusion, this strategy proved to be a powerful approach to purify a recombinant antibody and to ensure that the biological activity of the product was not impaired.#!/bin/bash # #./run.sh - run tests on a specific runner # # Usage: run.sh -f # # This script is in "setup_job" dir of the runner. if [[ "$#" -ne 2 ]]; then echo "usage:./run.sh " exit 1 fi if [[! -e "$PWD/build/linux" ]]; then echo "Can not find the build/linux/ folder!" exit 1 fi PROJECT_PATH=$1 BINARY=$2 LOG_FILE=$3 function do_test_on_runner() { ARGS=$@
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System Requirements:
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