Method for detecting ribosome inactivating proteins

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S091100, C536S023100, C536S024300, C536S024330, C536S025300

Reexamination Certificate

active

06864056

ABSTRACT:
The present invention concerns a method for detecting the presence of a ribosome inactivating protein in a sample. The method comprises contacting a sample suspected of containing a ribosome inactivating protein with a 2′-O-methylated oligonucleotide substrate having a GAxGA tetraloop wherein “Ax” is a deoxyribonucleoside comprising a fluorescent adenine derivative or analog base capable of emitting a fluorescence when released from the nucleoside, “Ax”, and detecting the presence of the fluorescent adenine derivative or analog base released from the tetraloop as an indication of the presence of the ribosome inactivating protein in the sample. The present invention is also directed to an assay kit and a reagent useful for carrying out the steps of the above method.

REFERENCES:
patent: 5538871 (1996-07-01), Nuovo et al.
patent: 6034233 (2000-03-01), Ecker et al.
Kao et al., Novel rapid method for the detection of ricin A-chain and related ribotoxins. Proceedings of the ERDEC Scientific Conference on Chemical and Biological Defense Research, pp. 483-491, on Nov. 17-20, 1998, MD, United States.*
Zamboni et al., High-pressure-liquid-chromatographic and fluorimetric methods for the determination of adenine released from ribosomes by ricin and gelonin. Biochem. J., 259, 639-643, 1989.*
Szewczak et al., The conformation of the sarcin/ricin loop from 28S ribosomal RNA. Proc. Natl. Acad. Sci. USA, 90, 9581-9585, Oct. 1993.*
Brigotti et al., A rapid and sensitive method to measure the enzymatic activity of ribosome-inactivating proteins. Nucleic Acids Res., 26, 4306-4307, 1998.

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