Epitope mapping and neuroprotective properties of a human single chain FV antibody that binds an internal epitope of amyloid-beta 1-42.
Active and passive immunotherapy targeted at the amyloid-beta (Abeta) peptide has been proposed as therapeutic approach against Alzheimer's disease (AD), and efforts towards the generation and application of antibody-based reagents that are capable of preventing and clearing amyloid aggregates are currently under active investigation. Previously, we selected and characterized a new anti-Abeta1-42 phage-displayed scFv antibody, designated clone b4.4, using a non-immune human scFv antibody library and demonstrated that a peptide based on the sequence of the Ig heavy chain (VH) complementarity-determining region (HCDR3) of this antibody fragment bound to Abeta1-42)and had neuroprotective potential against Abeta1-42 mediated neurotoxicity in rat hippocampal cultured neurons. In the present study, using novel computational methods and in vitro experiments we demonstrated that b4.4 binds to the central region of Abeta1-42. We also demonstrated that this scFv antibody binds to Abeta-derived diffusible ligands (ADDLs) and neutralizes the toxicity of both fibrillar and oligomeric forms of Abeta1-42 tested in vitro in SH-SY5Y cell cultures.
Solórzano-Vargas
R S
RS
Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, AP 70228, Cuidad Universitaria, México DF 04510, México.
Vasilevko
V
V
Acero
G
G
Ugen
K E
KE
Martinez
R
R
Govezensky
T
T
Vazquez-Ramirez
R
R
Kubli-Garfias
C
C
Cribbs
D H
DH
Manoutcharian
K
K
Gevorkian
G
G
D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000494:pharmacology*; D018604:Epitope Mapping; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000494:pharmacology*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D018696:Neuroprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D011485:Protein Binding
D016229:Amyloid beta-Peptides; D022801:Complementarity Determining Regions; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D018696:Neuroprotective Agents; D009842:Oligopeptides; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42); C055966:immunoglobulin Fv