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

doi:10.1007/s11745-997-0103-3

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 journal › Article › peer-review

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 language	English
Pages (from-to)	805-809
Number of pages	5
Journal	Lipids
Volume	32
Issue number	8
DOIs	https://doi.org/10.1007/s11745-997-0103-3
Publication status	Published - 1 Aug 1997

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title = "Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus",

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.",

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T1 - Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus

AU - Ziegler, Angelika

AU - Macintosh, Sybil M.

AU - Torrance, Lesley

AU - Simon, William

AU - Slabas, Antony R.

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

Abstract

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