A. Skerra, A GENERAL VECTOR, PASK84, FOR CLONING, BACTERIAL PRODUCTION, AND SINGLE-STEP PURIFICATION OF ANTIBODY F-AB FRAGMENTS, Gene, 141(1), 1994, pp. 79-84
The expression vector pASK84 was designed for the convenient cloning o
f immunoglobulin variable domain genes, as well as periplasmic secreti
on of the corresponding F-ab fragment in Escherichia coli. The plasmid
provides the constant domain genes of mouse IgG1/K with a hexa-histid
ine tag fused to the C terminus of the heavy chain. This strategy enab
les the rapid and efficient purification of the functional recombinant
F-ab fragment via immobilized metal affinity chromatography. The vers
atility of this expression and purification system is demonstrated usi
ng the variable domains of the well-characterized anti-lysozyme antibo
dy D1.3.