Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide

A. H. Tammer, G. Coia, R. Cappai, S. Fuller, C. L. Masters, P. Hudson, J. R. Underwood

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A recombinant Fab antibody, designated 1E8-4b, which reacts with the Alzheimer's disease (AD)-related Abeta peptides, Aβ[1-40], Aβ[1-42] and Aβ[1-43] has been developed. The 1E8-4b Fab was constructed by cloning the VHCH1 and VLCL domains from the parent hybridoma 1E8 antibody, reported previously to recognize these Aβ peptides. Briefly, a C-terminal Flag tag sequence was incorporated into this construct, which was ligated into the vector pHFA2 and expressed in Escherichia coli. Following purification on an M2 anti-Flag affinity column, the 1E8-4b recombinant Fab antibody was shown to bind plaques within sections of brain tissue from CERAD-defined AD patients by immunohistochemistry. ELISA, epitope mapping and immunoblotting confirmed the recognition of the Aβ[1-40/42/43] peptides by the 1E8-4b Fab. The 1E8-4b Fab did not recognize APP695 or APP770 which contain the Aβ sequence. The Aβ specificity of the recombinant 1E8-4b Fab antibody was identical to the parent 1E8 monoclonal antibody.
Original languageEnglish
Pages (from-to)453-463
Number of pages11
JournalClinical and Experimental Immunology
Volume129
Issue number3
DOIs
Publication statusPublished - 2002
Externally publishedYes

Keywords

  • recombinant Fab antibody
  • monoclonal antibody
  • Alzheimer’s disease
  • Aβ peptide

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