A new ELISA for quantification of brassinosteroids in plants

• Conjugates of 28-norcastasterone with HRP and BSA were prepared.
• Polyclonal antibodies to 28-norcastasterone were isolated from rabbit blood.
• New enzyme immunoassay for quantification of 6-oxo-brassinosteroids was elaborated.
• The direct method of BS enzyme immunodetection was proposed.
• The selective determination of individual hormones of the 28-homo-BS was carried out.

Starting from (22R,23R)-2α,3α,22,23,26-pentahydroxy-5α-cholestan-6-one 26-hemisuccinate, conjugates of 28-norcastasterone with horse radish peroxidase and bovine serum albumin were prepared. The latter conjugate was injected into rabbits; produced polyclonal antibodies were used to quantitate 6-keto-brassinosteroids. The newly developed analytical system was used in combination with two other immunoenzymatic assays for brassinosteroids to determine individual compounds of this series. In addition, a direct method of brassinosteroid analysis was proposed. It has the advantage of requiring no sample pretreatment steps such as extraction with organic solvents and chromatography.

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