Single chain Fv antibodies directed against the 37 kDa/67 kDa laminin receptor as therapeutic tools in prion diseases.
Transmissible spongiform encephalopathies are a group of neurological disorders associated with the deposition of PrP(Sc), an abnormal form of the cellular prion protein PrP(c). The 37 kDa/67 kDa laminin receptor (LRP/LR) has been identified as a prion receptor and several lines of evidence strongly suggest that this protein plays a role during prion pathogenesis. Here we report the selection of recombinant single chain antibodies (scFvs) directed against LRP from naïve and synthetic phage scFv libraries for therapeutic application. Western blotting and FACS analysis confirmed a specific LRP/LR recognition pattern of the two selected scFvs S18 and N3. Both scFvs specifically interfered with the PrP/LRP interaction in vitro. High yield production of the scFvs of approx. 1mg/l of culture medium was achieved in E. coli. Passive immunotransfer of the scFv S18 antibody reduced PrP(Sc) levels by approx. 40% in the spleen of scrapie infected C57BL/6 mice 90 days post scFv injection, suggesting that scFv S18 interferes with peripheral PrP(Sc) propagation, without a significant prolongation of incubation and survival times.
Zuber
Chantal
C
Laboratorium für Molekulare Biologie-Genzentrum, Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
Knackmuss
Stefan
S
Rey
Clémence
C
Reusch
Uwe
U
Röttgen
Peter
P
Fröhlich
Thomas
T
Arnold
Georg J
GJ
Pace
Claudia
C
Mitteregger
Gerda
G
Kretzschmar
Hans A
HA
Little
Melvyn
M
Weiss
Stefan
S
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000494:pharmacology*; D000918:Antibody Specificity / Q000187:drug effects / Q000276:immunology; D004195:Disease Models, Animal; D018604:Epitope Mapping; D005260:Female; D006801:Humans; D007116:Immunization, Passive; D007135:Immunoglobulin Variable Region / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018621:PrPC Proteins / Q000378:metabolism; D018620:PrPSc Proteins / Q000378:metabolism; D017096:Prion Diseases / Q000628:therapy*; D011485:Protein Binding / Q000187:drug effects; D017474:Receptors, Laminin / Q000037:antagonists & inhibitors*; D012608:Scrapie / Q000378:metabolism; D013154:Spleen / Q000187:drug effects / Q000378:metabolism / Q000473:pathology
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D018621:PrPC Proteins; D018620:PrPSc Proteins; D017474:Receptors, Laminin