Generation of recombinant antibody fragments for membrane protein crystallization

24.03.2015

Mir SH, Escher C, Kao WC, Birth D, Wirth C, Hunte C.

Methods Enzymol. 2015;557:201-18

Methods Enzymol.            online article

Membrane proteins are challenging targets for crystallization and structure determination by X-ray crystallography. Hurdles can be overcome by antibody-mediated crystallization. More than 25 unique structures of membrane protein:antibody complexes have already been determined, most of them with hybridoma-derived antibody fragments either in Fab or Fv fragment format. The publication summarizes background and current status of the strategy and describes in detail current protocols of well-established methods for the immunization, the selection, and the characterization of antibodies, as well as the cloning, the production, and the purification of recombinant antibodies useful for structural analysis of membrane proteins.