Acridinium ester labelling and purification of nucleotide probes

Details Acridinium ester labelling and purification of nucleotide probes Acridinium ester labelling and purification of nucleotide probes

1988-12-12

1993-02-09

Rollins, John W.

Organic compounds -- part of the class 532-570 series

Organic compounds

Carbohydrates or derivatives

435 6, C07H 1700, C12Q 168

Patent

active

051854392

ABSTRACT:
Methods for the construction, labelling and subsequent purification of nucleic acid probes containing primary amines with acridinium esters 4-(2-succinimidyloxycarbonyl-ethyl)phenyl-10-methylacridinium-9-carboxylat e fluorosulfonate). The method for attaching acridinium esters to probes uses high (0.1 to 50 mM) acridinium ester concentrations achieved using organic solvent in concentrations of 20% to 80% by volume, and may be carried out either in solution, or with one or the other of the acridinium ester or the probe suspended in solution. Purification (the separation of labelled probe from unlabelled probe and free label) involves (1) first removing most of the free acridinium ester label from probe using rapid separation techniques, then (2) removing substantially all remaining free label from the probe and separating labelled probe from unlabelled probe, involves specific applications of ion exchange, reverse phase or hydroxyapatite HPLC.

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Septak, the Chemical Abstracts, 107:39648V (1987).
Hart et al., the Chemical Abstracts 107:232557t (1987).
Fieser Experiments in Organic Chemistry (1957) pp. 281-294 H. Heath & Co., U.S.
Weeks et al "Acridinium Esters as High-Specific-Activity Labels in Immunassay" Clin. Chem. vol. 29, #8, 1983, pp. 1474-1479.

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