Amersham Pharmacia Biotech UK Limited

Ge Healthcare UK Limited

Nycomed Amersham Plc

Adrian Christopher Simmonds

David Loakes

Alan L Hamilton

Clifford Leslie Smith

Daniel M Brown

Shiv R Kumar

Mark McDougall

Satyam Nampalli

Fergal Hill

Mark Samuel Briggs

Ian Edward Bruce

Elizabeth Swann

Christopher John Moody

James N Miller

Karen Williams

William Jonathan Cummins

Raj Odedra

David Williams

Alison Claire Sweet

Patrick Finn

Susan Janet Fowler

Lee Pickering

Timothy Stone

A matched set of fluorescent dyes is provided, wherein each dye of the set is capable of covalent attachment to a protein and wherein each of the dyes has a molecular structure and a charge that is matched one with the other, such that relative electrophoretic mobility of a protein labeled with one dye of the set is the same as the electrophoretic mobility of the protein labeled with a different dye of the set. The matched set comprises at least two different fluorescent dyes of formula: wherein n is 1, 2, or 3; Zand Zindependently represent the carbon atoms necessary to complete a phenyl or naphthyl ring system; one of groups Rand Ris a target bonding group; remaining group Ror Ris selected from &#x2014;(CH)&#x2014;W or &#x2014;(CH)&#x2014;H; group Ris hydrogen, except when either Ror Ris &#x2014;(CH)&#x2014;H, in which case Ris W; and W is selected from sulphonic acid and sulphonate. The invention also provides a method for saturation labeling of a protein with a fluorescent dye so as to label all available target amino acid, suitably cysteine, residues in the protein, thereby giving a single population of labeled protein molecules.

Reagents and a method for saturation labelling of proteins

Nucleotides comprising a reporter moiety and a polymerase enzyme blocking moiety in which the reporter moiety does not also act as a polymerase enzyme blocking moiety are described. Also described are compounds of Formula (I): wherein W is a phosphate group, B is a base, Y is a linker comprising an enzyme-cleavable group, Ris a reporter moiety, Ris selected from H or OH, Z and Z' are selected from H, OH, or a group X&#x2014;R, wherein X is a linker comprising an enzyme-cleavable group and Ris a polymerase enzyme blocking group, provided that at least one of Z and Z&#x2032; is X&#x2014;R.

Nucleotide analogues comprising a reporter moiety and a polymerase enzyme blocking moiety

Nucleoside analogues or base analogues having structure (I), where X&equals;O or NH or S, Y&equals;N or CHR or CR , W&equals;N or NR or CHR or CR or S, n&equals;1 or 2; each R is independently H or O or alkyl or alkenyl or alkoxy or aryl or a reporter moiety; where necessary (i.e. when Y and/or W is N or CR where R is not O) a double bond is present between Y and W or W and W, and Q is H or a sugar or a sugar analogue or a nucleic acid backbone or backbone analogue.

Base analogues

Nucleoside analogues have structure (2) wherein Q is H or a sugar moiety or sugar analogue or a modified sugar or a nucleic backbone or backbone analogue, W is an alkylene or alkenylene chain of 0-5 carbon atoms, any of which may carry a substituent R , X is O or N or NR or CR , X&prime; is O or S or N, provided that when X&prime; is O or S, then X is C, Y is CH or N, R is NH or SMe or SO Me or NHNH , each of R and R is independently H or F or alkyl or alkenyl or aryl or acyl or a reporter moiety, R is independently H or alkyl or alkenyl or aryl or acyl or a reporter moiety, and R is H or &boxH;O or F or alkyl or aryl or a reporter moiety.

Tricyclic base analogues

Nucleoside analogues containing the degenerate base analogue P and derivatives thereof are provided with reporter moieties preferably comprising signal moieties. The nucleoside analogues are useful for labeling DNA or RNA or for incorporating in oligonucleotides.

Nucleoside analogues

Nucleoside analogues in which a group M replaces the natural base where M is (1) or (2) or (3), where each of X,, X,and X,are C or N, each of R,and R,is the same or different and each is H, NO,, CO, COR,, OR,, CN, O, CON(R,),, COOR,, SO,R,, SO,R,, SR,, NHCHO, (CH,),(R,),, halogen, or a reporter moiety, each of R,and R,is H or hydrocarbyl or a reporter moiety, and n is 0-4. The analogues are substrates for polymerase and terminal transferase enzymes.

Benzophenoxazine dyes

Growing community of inventors

Amersham, United Kingdom

Adrian Christopher Simmonds