Purification of recombinant S-layer fusion proteins in nanobiotechnology
Abstract
Crystalline bacterial cell surface layers (S-layers) represent the outermost cell envelope component of many bacteria and archaea. S-layers are composed of an identical protein or glycoprotein species with the ability to self-assemble into lattices exhibiting either oblique, square or hexagonal symmetry. Because of the specific properties of S-layer lattices, such as the high density of functional groups on the outermost surface and in the pore areas, the presence of pores identical in size and morphology and the ability of isolated S-layer subunits to recrystallize into closed monolayers on solid supports, lipid films and liposomes, S-layer proteins can be considered as new biomaterial with broad application potential in molecular nanotechnology and nanobiotechnology. S-layers are currently used as matrix for the controlled immobilization of biologically active macromolecules for the production of S-layer-based dipstick-style solid-phase immunoassays as well as for the development of biosensors, biochips and complex biomimetic immunogens. Moreover, S-layer lattices were found to increase the thermal and mechanical stability of unilamellar liposomes and Langmuir films. Up to now, S-layer proteins have been isolated from various wild-type strains, but in future developments, functionalized S-layer fusion proteins produced by recombinant technologies shall be used: the part of the protein sequence not involved in the self-assembly process shall be replaced by specific functions, e. g. an immunoglobulin-binding domain, which implies that functionalized S-layer fusion proteins are still capable to self-assemble and to recrystallize into closed monolayers. Moreover, the specific function shall be exposed on the S-layer surface or within the pores to be accessible for further binding and reaction partners. For preparing them with a purity higher than 98 %, an appropriate purification procedure has to be developed.
Project staff
Seta Küpcü
Assistenzprofessor Dipl.-Ing.Dr.techn. Seta Küpcü
seta.kuepcue@boku.ac.at
Tel: +43 1 47654-80456
Project Staff
01.07.2000 - 30.06.2001
Uwe B. Sleytr
em.o.Univ.Prof. Dipl.-Ing.Dr. Uwe B. Sleytr
uwe.sleytr@boku.ac.at
Project Staff
01.07.2000 - 30.06.2001