A General Assay for Antibody Catalysis Using Acridone as a Fluorescent Tag

A simple and highly sensitive catalysis assay is demonstrated based on analyzing reactions with acridone-tagged compounds by thin-layer chromatography. As little was 1 pmol of product is readily visualized by its blue fluorescence under UV illumination and identified by its retention factor (Rf). Ea...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1996-04, Vol.93 (9), p.4251-4256
Main Authors: Reymond, Jean-Louis, Koch, Thomas, Schröer, Josef, Tierney, Emily
Format: Article
Language:English
Subjects:
Acridines - chemical synthesis
Acridines - chemistry
Alcohol Dehydrogenase - metabolism
Alcohols
Aldehydes
Alkenes
Animals
Antibodies
Catalysis
Catalytic antibodies
Chemistry
Chromatography
Chromatography, Thin Layer - methods
Epoxy Compounds
Fluorescence
Fluorescent Dyes
Horses
Hydrazides
Hydrazones
Immunity (Disease)
Indicators and Reagents
Isomers
Magnetic Resonance Spectroscopy
Oxidation-Reduction
Sensitivity and Specificity
Spectrometry, Fluorescence
Stereoisomerism
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Summary:A simple and highly sensitive catalysis assay is demonstrated based on analyzing reactions with acridone-tagged compounds by thin-layer chromatography. As little was 1 pmol of product is readily visualized by its blue fluorescence under UV illumination and identified by its retention factor (Rf). Each assay requires only 10 μ l of solution. The method is reliable, inexpensive, versatile, and immediately applicable in repetitive format for screening catalytic antibody libraries. Three examples are presented: (i) the epoxidation of acridone labeled (S)-citronellol. The pair of stereoisomeric epoxides formed is resolved on the plate, which provides a direct selection method for enantioselective epoxidation catalysts. (ii) Oxidation of acridone-labeled 1-hexanol to 1-hexanal. The activity of horse liver alcohol dehydrogenase is detected. (iii) Indirect product labeling of released aldehyde groups by hydrazone formation with an acridone-labeled hydrazide. Activity of catalytic antibodies for hydrolysis of enol ethers is detected.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.93.9.4251