Rapid Analysis of Small Samples Containing Forskolin Using Monoclonal Antibodies

Yanagihara, H.; Sakata, Ryuji; Shoyama, Yukihiro; Murakami, Hiroki

doi:10.1055/s-2006-957844

Cited by 29 publications

(11 citation statements)

References 6 publications

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“…In order to confirm the antigen conjugate formation, mice splenocytes immunized with the antigen conjugate of 7 and myeloma cells were fused to make hybridoma, by the polyethylene glycol method. Anti-forskolin antibody formation was confirmed by enzyme linked immuno-sorbent assay (ELISA) as in a previous report [18,19]. Anti-forskolin antibodies appear to be of satisfactory affinity and specificity.…”

Section: Resultssupporting

confidence: 73%

Direct determination of naturally occurring biologically active compound–serum albumin conjugate by matrix‐assisted laser desorption/ionization mass spectrometry

Tanaka

Morimoto

et al. 2000

Journal of Spectroscopy

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Opium alkaloids were conjugated with bovine serum albumin (BSA) to give individual antigen conjugates which were analyzed using BSA as an internal standard by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. It became clear that 9 molecules of thebaine were conjugated in a thebaine–BSA conjugate. Codeine and morphine contents in individual conjugates were determined to be 12 and 6 molecules, respectively. Using a relative molecular mass of BSA, 11.5 molecules of forskolin conjugated with BSA. Tetrahydrocannabinolic acid (THCA)–carrier protein conjugates usingβ-alanine or withoutβ-alanine as a linker were analyzed. From these results it became evident that the hapten molecules of THCA–β-alanine–BSA, THCA–BSA, THCA–β-alanine–human serum albumin (HSA) and THCA–HSA were 17.2, 12.7, 27.8 and 54.4, respectively. Crocin–HSA conjugates were synthesized by two methods using succinate and NaIO4cleavage of sugar moiety. A broad molecular peak of crocin–HSA conjugate appeared at aroundm/z98,342 resulting in 30 molecules of crocin-hemisuccinate conjugated with HSA. The crocin–HSA synthesized via NaIO4oxidation showed a sharp molecular peak around 69,504 compared to that of hemisuccinate preparation. The number of crocin combined with HSA was, however, smaller 3.1 molecules.Analysis of monoclonal antibody by MALDI mass spectrometry determined the exact molecular weight and their purities.

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Section: Resultssupporting

confidence: 73%

Direct determination of naturally occurring biologically active compound–serum albumin conjugate by matrix‐assisted laser desorption/ionization mass spectrometry

Tanaka

Morimoto

et al. 2000

Journal of Spectroscopy

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“…Therefore, the leaves are thought to be the primary location for forskolin synthesis. We previously reported the forskolin concentration in clonally propagated plant organs of C. forskohlii [24]. Tuberous roots and the stem base were determined to contain a higher concentration of forskolin than the organs.…”

Section: Resultsmentioning

confidence: 99%

Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohliiBriq

et al. 2004

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Background: Isopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq.

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“…One recently developed method to answer the request is an immunoassay system [10][11][12][13][14] in which specific monoclonal antibodies (MAbs) against small molecular weight compounds are prepared and, by using the specific affinity of the MAbs to antigen, reliable quantitative and qualitative analyses of those compounds can be performed.…”

Section: Introductionmentioning

confidence: 99%

Production of monoclonal antibodies against antitumor cyclohexapeptide RA-VII from Rubia cordifolia and their characterization

2011

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An immunoassay system was established for the estimation of the quantity of an antitumor cyclic hexapeptide RA-VII (1) from Rubia cordifolia L. and R. akane Nakai (Rubiaceae). First, 1 was converted into its hapten, which was then conjugated with a carrier protein to be used as an effective antigen to obtain its monoclonal antibody (MAb). In the resulting conjugate, the molecular ratio between 1 and the carrier protein as assayed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was about 5:1. Then, the splenocytes from the mouse immunized with the conjugate were fused with mouse myeloma cells to produce hybridoma, secreting MAb against 1. Two clones were isolated, one producing MAb IgG(1) and the other IgM, both having a κ light chain. The sensitivity and cross-reactivity of the thus obtained MAb were also assayed.

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Rapid Analysis of Small Samples Containing Forskolin Using Monoclonal Antibodies

Cited by 29 publications

References 6 publications

Direct determination of naturally occurring biologically active compound–serum albumin conjugate by matrix‐assisted laser desorption/ionization mass spectrometry

Direct determination of naturally occurring biologically active compound–serum albumin conjugate by matrix‐assisted laser desorption/ionization mass spectrometry

Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohliiBriq

Production of monoclonal antibodies against antitumor cyclohexapeptide RA-VII from Rubia cordifolia and their characterization

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