Hi Ivan, Just have a look at this paper:
Skerra, A. (1994) A general vector, pASK84, for cloning, bacterial production, and single-step purification of antibody Fab fragments. Gene 141, 79-84. We (and other labs) have used this plasmid as well as its successors for the cloning and preparation of numerous recombinant Fabs, and a couple of them have also been crystallized successfully (some X-ray structures still awaiting publication): Schiweck, W. & Skerra, A. (1995) Fermenter production of an artificial Fab fragment, rationally designed for the antigen cystatin, and its optimized crystallization through constant domain shuffling. Proteins: Struct. Funct. Genet. 23, 561-565. I will be happy to share these plasmids for academic research purposes. Best regards, Arne Skerra Am 18.11.2014 um 19:39 schrieb xaravich ivan <[email protected]>: > Hi everyone, > I have a few single chain antibodies (scFV) that I would like to express in a > Fab format in bacteria for crystallization purposes. > > Could you suggest some plasmids that have success in such kind of projects? > Are there commercial plasmids consisting of antibody constant regions ready > for bacterial expression, where I can just clone my scFv and it would be > ready for Fab expression (probably periplasmic or secreted in supernatant) > > If commercial plasmids are not available, could you suggest where I could > request them from? > > Thanks in advance, > Ivan Prof. Dr. Arne Skerra Lehrstuhl f. Biologische Chemie | Technische Universitaet Muenchen Emil-Erlenmeyer-Forum 5 | 85350 Freising-Weihenstephan | Germany Phone: +49 (0)8161 71-4351 | Fax: -4352 eMail: [email protected] | http://www.wzw.tum.de/bc
