Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1996-02-05
1998-06-23
Marschel, Ardin H.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 221, 536 231, 536 243, 536 253, 536 2531, 536 2532, C12Q 168, C07H 1900, C07H 2102, C07H 2104
Patent
active
057703671
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 of PCT/GB34/01675 filed on Aug. 1, 1994.
In biological and chemical analyses, the use of analyte molecules labelled with reporter groups is routine. This invention addresses the idea of providing reagents having at least two analyte groups linked to one or more reporter groups. Such reagents can be used, in ways described below, to generate much more analytical information than can simple labelled analytes. It is possible to code reporter groups so that reagents carrying multiple analyte groups and multiple reporter groups can by synthesised combinatorially and used simultaneously and the reporter groups resolved in the analytical stage.
WO 93/06121 (Affymax) describes a synthetic oligomer library comprising a plurality of different members, each member comprising an oligomer composed of a sequence of monomers linked to one or more identifier tags identifying the sequence of monomers in the oligomer. The linkage between the oligomer and the identifier tag preferably comprises a solid particle. The identifier tag is preferably an oligonucleotide.
Proc. Natl. Acad Sci., Vol 89, No. 12, 15 Jun. 1992, pages 5381-5383 (S Brenner and R A Lerner) describe encoded combinatorial chemistry for making a library of reagents each containing a genetic oligonucleotide tag.
In Rapid Communications in Mass Spectrometry, Vol 6, pages 369-372 (1992), G R Parr et al describe matrix-assisted laser desorption/ionisation mass spectrometry of synthetic oligodeoxyribonucleotides.
In Nucleic Acids Research, Vol 21, No. 15, 25 July 1993, pages3347-3357, E Nordhoff et al describe the ion stability of nucleic acids in infra-red matrix-assisted laser desorption/ionisation mass spectrometry. to which the analyte moiety and the tag moiety are both attached. Preferably the analyte moiety is a chain of n analyte residues, and the tag moiety is a chain of up to n reporter groups, the reporter group at each position of the tag chain being chosen to designate the analyte residue at a corresponding position of the analyte chain. n is an integer of at least 2, preferably 3 to 20.
The invention may be used for the detection of all analytes of interest. These include, but are not limited to, a protein/peptide chain so that the analyte residues are amino acid residues; a nucleic acid/oligonucleotide chain so that the analyte residues are nucleotide residues; a carbohydrate chain so that the analyte residues are sugar residues. Additionally the analyte may be a class of small molecules with biological, pharmacological or therapeutic activity. For example it could be a core molecule with the ability to vary various substituent groups eg. alkyl, esters, amines, ethers etc in a combinatorial manner with mass spectrometry tags.
The tag moiety and/or the or each reporter group in it is capable of being observed/detected/analysed so as to provide information about the nature of the analyte moiety, and/or the analyte residues in it.
In one embodiment, the reagent has the formula A--L--R where A is a chain of n analyte residues constituting the analyte moiety, L is the linker, R is a chain of up to n reporter groups constituting the tag moiety, and n is 2-20, wherein the tag moiety contains information defining the location of analyte residues in the analyte moiety.
The tag moiety consists of one or more reporter groups distinguishable by mass and thus capable of being analysed by mass spectrometry. The reporter groups may be chemically different and thus distinguished from one another by molecular weight. Or the reporter groups may be chemically identical, but distinguished from one another by containing different isotopes (e.g. .sup.12 C/.sup.13 C and .sup.1 H/.sup.2 H as discussed below). The tag moiety is, and/or the reporter groups are, suitable or adapted for analysis by mass spectrometry e.g. after cleavage by photochemical or other means from the reagent.
The advantages of mass spectrometry as a detection system are: its great sensitivity--only a few hundred molecules are needed to give a good signal; its wide dynamic rang
REFERENCES:
patent: 5258506 (1993-11-01), Urdea et al.
S. Brenner et al., Proc. Natl. Acad. Sci., 89(12), 5381-5383 (Jun. 15, 1992).
G.R. Parr et al., Rapid Communication in Mass Spectrometry, 6(6), 369-372 (Jun. 1992).
E. Nordhoff et al., Nucl. Acids Res., 21(15), 3347-3357 (Jul. 25, 1993).
Cummins William Jonathan
Southern Edwin
Marschel Ardin H.
Oxford Gene Technology Limited
Riley Jezia
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