Production of monoclonal antibodies to a steroid plant growth regulator

Abstract

1984) Production of monoclonal antibodies to a steroid plant growth regulator.Can. J . Biochem. Cell Biol. 62, [715][716][717][718][719][720][721] The active component of the brassin complex of higher plants has been shown to be a C28 steroidal lactone called brassinolide. This compound has been shown to enhance both cell division and cell growth. Monoclonal antibodies directed against a synthetic brassinosteroid have been produced in CAFB mice. The mice were immunized with a saline solution containing the steroid hapten noncovalently bound to chloroform-methanol extracted fetal calf serum. Hybridomas were generated by fusing primed mouse spleen cells with SP2/0 myeloma cells. Sixty-one hybridoma clones were screened; four of these gave positive enzyme-linked immunosorbent assay (ELISA) values with the brassinosteroid hapten. One of these clones produced an immunoglobulin G antibody with a high specificity for the synthetic antigen. The brassinosteroid hapten which differs from the natural brassinolide by the position of a methyl group on the C24 carbon gave a considerably higher ELISA value than the natural growth regulator and several other plant steroids. The distribution of brassinolide in Brassica napus tissues was examined by ELISA with the monoclonal antibody.Horgen, P. A., Nakagawa, C. W. & b i n , R. T. (1984) Production of monoclonal antibodies to a steroid plant growth regulator.Can. J . Biochem. Cell Biol. 62, 715-721 Le constituant actif du complexe brassin des plantes sup6rieures est une lactone stCroi'de comportant 28 atomes de carbsne, appelCe brassinolide. Ce compose favorise la division cellulaire et la croissance cellulaire. Nous avons produit des anticorps monoclonaux dirigCs contre un brassinsstCroi'de synthCtique dans les souris CAF,. Les souris Ctaient immunisCes avec une solution saline contenant 19hapt&ne sttrside lit de fason non covalente au serum de veau foetal extrait avec un melange de chlorofome-mCthano1. Des hybridomes sont gtndrts par fusion de cellules de rates de souris immunisdes avec les cellules du mytlome SP2/0. Nous avons examine 61 clones d9hybridomes; quatre de ces clones donne des valeurs ELZSA (test immunoenzymatique) positives avec l'hapdne brassinostQoide. L'un de ces clones produit un anticorps immunoglobuline C avec une spCcificitC ClevCe pour l'antigkne synthetique. L'hapthe brassinosdroi'de, qui diff&re du brassinolide naturel par la presence d'un groupe mkthyle sur le carbsne 24, donne une valeur ELISA considCrablement plus Clevie que le rCgulateur naturel de la croissance et plusieurs autres stCroi'des vtgetaux. Nous avons examine la distribution du brassinolide dans les tissus de Brassica napus par l'essai ELISA avec l'anticorps monoclonal.[Traduit par ]la revue]

Brassinosteroid phytohormones: structure, bioactivity and applications

“…The biological detection of brassinosteroids was initially performed by the bean second internode bioassay, a test gradually substituted by the rice lamina inclination bioassay (Wada et al, , 1984Takeno and Pharis, 1982;Kim et al, 1990) and the wheat leaf unrolling bioassay (Wada et al, 1985;Takatsuto, 1994), while the immunological methods are less used (Horgen et al, 1984;Yokota et al, 1990;Schlagenhaufer et al, 1991;Taylor et al, 1993).…”

A New Type of Modified Brassinosteroids for Enzyme-linked Immunosorbent Assay

“…A good alternative to determine steroids in natural sources is immunochemical analysis requiring much simpler sample preparation procedures. Several attempts have been made on the development of immunochemical assays for BS analysis [22][23][24], which were mostly based on radio-immune methodology using radio-labeled BS as markers in an immuno-competitive assay. Until recently there were no published results on the development of an immunoenzymatic approach to BS-analysis, where an enzyme (usually HRP) is used for the labeling of BS.…”

Amino Acid Catalyzed Neogenesis of Carbohydrates: A Plausible Ancient Transformation

“…For the immunoassays, the hydroxyl or 6‐ketone is usually utilized as modification sites for the further conjugation with the carrier protein. Horgen and co‐workers7 first developed monoclonal antibodies using a brassinolide analogue conjugate, but the high cross‐reactivity with other major plant sterols, such as sitosterol and ergosterol, made it unfeasible for the analysis of BRs. Yokota and co‐workers8 synthesized hapten using castasteronecarboxymethoxylamine oxime conjugated to bovine serum albumin (BSA), and then produced anti‐serum with affinity to some naturally occurring brassinosteroids.…”