Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus

Angelika Ziegler*, Sybil M. Macintosh, Lesley Torrance, William Simon, Antony R. Slabas

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single-chain F(v) (scF(v)), displayed on filamentous phage. Analysis of the selected clones by BstNI fingerprinting and nucleotide sequencing showed that the scF(v) were derived from three different human V(H) germline genes. The binding specificities were confirmed by Western blots and ELISA. The scF(v) preparations reacted with B. napus ENR, but not with β-keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scF(v) 3.13 recognizes an epitope located within the N-terminal 80 amino acids of the enzyme molecule. The scF(v) were used to detect ENR directly in extracts of B. napus seeds.

Original languageEnglish
Pages (from-to)805-809
Number of pages5
JournalLipids
Volume32
Issue number8
DOIs
Publication statusPublished - 1 Aug 1997

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