Magnetizable antibody-like proteins.
Use of paramagnetic particles to isolate molecules or cells from complex media is well established. Typically, particles are manufactured and coated with a biological molecule that confers specific biorecognition. Incubation of particles with sample and exposure to magnetic fields isolates the species of interest. We have designed, produced and assessed magnetized fusion proteins consisting of the antigen-binding portion of an antibody (single chain variable fraction; scFv) fused to the heavy chain of the iron-binding protein ferritin. The fusion protein subunits expressed in E. coli assemble to form a fusion protein consisting of a ferritin sphere with scFvs on the surface. The fusion proteins were chemically magnetized by introducing a paramagnetic iron core. The resultant fusion protein was shown to be magnetizable and capable of binding target antigens. These "organic" magnetizable particles possess a number of theoretical advantages over traditional inorganic particles.
Dehal
Prabhjyot Kaur
PK
Research and Development, Axis-Shield Diagnostics Ltd, The Technology Park, Dundee, UK. Prabhjyot.Dehal@axis-shield.com
Livingston
Christine Frances
CF
Dunn
Claire Geekie
CG
Buick
Richard
R
Luxton
Richard
R
Pritchard
David John
DJ
D000595:Amino Acid Sequence; D000818:Animals; D001709:Biotechnology / Q000379:methods; D001792:Blood Platelets / Q000378:metabolism; D005293:Ferritins / Q000737:chemistry* / Q000235:genetics; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics; D013052:Spectrometry, X-Ray Emission; D020349:Surface Plasmon Resonance
D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D005293:Ferritins; D052203:Ferrosoferric Oxide