A general vector, pASK84, for cloning, bacterial production, and single-step 
purification of antibody Fab fragments

Skerra, Arne

doi:10.1016/0378-1119(94)90131-7

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A general vector, pASK84, for cloning, bacterial production, and single-step purification of antibody F_ab fragments

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Skerra, Arne
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Skerra, A. (1994). A general vector, pASK84, for cloning, bacterial production, and single-step purification of antibody F_ab fragments. Gene, 141(1), 79-84. doi:10.1016/0378-1119(94)90131-7.

Cite as: https://hdl.handle.net/21.11116/0000-0007-AA60-D

Abstract

The expression vector pASK84 was designed for the convenient cloning of immunoglobulin variable domain genes, as well as periplasmic secretion of the corresponding F_a fragment in Escherichia coli. The plasmid provides the constant domain genes of mouse IgG1/kappa with a hexa-histidine tag fused to the C terminus of the heavy chain. This strategy enables the rapid and efficient purification of the functional recombinant F_a fragment via immobilized metal affinity chromatography. The versatility of this expression and purification system is demonstrated using the variable domains of the well-characterized anti-lysozyme antibody D1.3.