RECOMBINANT ANTIBODY FRAGMENTS THAT DETECT ENOYL ACYL CARRIER PROTEINREDUCTASE IN BRASSICA-NAPUS

Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragmen ts, single-chain F-v (scF(v)), displayed on filamentous phage. Analysi s of the selected clones by BstNI fingerprinting and nucleotide sequen cing showed that the scF(v) were derived from three different human VH germline genes. The binding specificities were confirmed by Western b lots and ELISA. The scF(v) preparations reacted with B. napus ENR, but not with beta-keto reductase, nor enoyl reductase from Escherichia co il. Analysis of fragments generated by CNBr treatment indicates that t he 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.