Germline humanization of a murine Abeta antibody and crystal structure of the humanized recombinant Fab fragment.
Alzheimer's disease is the most common form of dementia, affecting 26 million people worldwide. The Abeta peptide (39-43 amino acids) derived from the proteolytic cleavage of the amyloid precursor protein is one of the main constituents of amyloid plaques associated with disease pathogenesis and therefore a validated target for therapy. Recently, we characterized antibody fragments (Fab and scFvs) derived from the murine monoclonal antibody WO-2, which bind the immunodominant epitope ((3)EFRH(6)) in the Abeta peptide at the N-terminus. In vitro, these fragments are able to inhibit fibril formation, disaggregate preformed amyloid fibrils, and protect neuroblastoma cells against oligomer-mediated toxicity. In this study, we describe the humanization of WO-2 using complementary determining region loop grafting onto the human germline gene and the determination of the three-dimensional structure by X-ray crystallography. This humanized version retains a high affinity for the Abeta peptide and therefore is a potential candidate for passive immunotherapy of Alzheimer's disease.
Robert
Remy
R
CSIRO Molecular and Health Technologies, 343 Royal Parade, Parkville, Victoria 3052, Australia. remy.robert@csiro.au
Streltsov
Victor A
VA
Newman
Janet
J
Pearce
Lesley A
LA
Wark
Kim L
KL
Dolezal
Olan
O
D015502:Absorptiometry, Photon; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D005854:Germ Cells / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D016415:Sequence Alignment
D016229:Amyloid beta-Peptides; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins