Immunoaffinity columns for isolation of abscisic acid in conifer seedlings.

Spleen cells of mice immunized with cis,trans (±) abscisic acid (ABA) conjugated to bovine serum albumin (BSA) were fused with myeloma cells to produce hybridomas. Hybridomas secreting antibodies to cis,trans (+) ABA were detected by enzyme-linked immunoassay and injected into the peritoneal cavity of mice to produce ascites fluid. Antibodies purified by size-exclusion chromatography were linked to cyanogen bromide-activated Sepharose beads and used to prepare immunoaffinity columns (IAC). Semi-purified conifer seedling {Douglas-fir [Pseudotsuga menziesii (Mirb) Franco] lodgepole pine Pinus contorta Dougl), cedar (Thuja plicata D. Don), and spruce [Picea glauca (Moench) Voss)]} extracts were passed through IAC, washed thoroughly with water and then eluted with methanol. ABA in the methanol eluate was analysed by high performance liquid chromatography with UV detection. UV absorption of extracts before and after purification through IAC indicated an excellent purification of extract by IAC. The identity of the ABA peak was confirmed by gas chromatography-mass spectrometry. [ABSTRACT FROM AUTHOR]