Labeling reagents, methods for synthesizing such reagents and methods for detecting biological molecules
申请人:BIO MERIEUX
公开号:US20030186448A1
公开(公告)日:2003-10-02
The present invention relates to a temperature-stable labeling reagent of formula:
1
in which
R
1
represents H or an alkyl, aryl or substituted aryl group,
R
2
represents a detectable marker or at least two detectable markers linked together by at least one multimeric structure,
L is a linking arm containing a linear succession of at least two covalent bonds and n an integer equal to 0 or 1,
R
3
and R
4
represent independently of each other: H, NO
2
, Cl, Br, F, I, R
2
-(L)
n
-Y—X—, OR, SR, NR
2
, R, NHCOR, CONHR, COOR with R=alkyl or aryl,
A is a linking arm containing at least one covalent double bond allowing conjugation of the diazo functional group with the aromatic ring and u is an integer between 0 and 2, preferably equal to 0 or 1, and
—Y—X— represents —CONH—, —NHCO—, —CH
2
O—, —CH
2
S—.
The present invention also describes a method for synthesizing said markers as well as applications for labeling biological molecules, in particular nucleic acids, with a labeling reagent carrying the diazomethyl functional group.
The invention finds a preferred application in the field of diagnosis.
本发明涉及一种公式为1的温度稳定标记试剂:
其中,R1表示H或烷基、芳基或取代芳基基团,R2表示可检测的标记或至少两个通过至少一个多聚物结构连接在一起的可检测标记,L是含有至少两个共价键的线性链连接臂,n是等于0或1的整数,R3和R4分别独立地表示:H、NO2、Cl、Br、F、I、R2-(L)n-Y-X-、OR、SR、NR2、R、NHCOR、CONHR、COOR,其中R=烷基或芳基,A是含有至少一个共价双键的连接臂,允许将重氮甲基官能团与芳香环共轭,u是介于0和2之间的整数,优选等于0或1,而-Y-X-表示-CONH-、-NHCO-、-CH2O-或-CH2S-。
本发明还描述了一种合成所述标记的方法,以及使用携带重氮甲基官能团的标记试剂标记生物分子,特别是核酸的应用。
本发明在诊断领域中找到了一种首选应用。