Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
1998-05-28
2001-03-06
Marschel, Ardin H. (Department: 1631)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S022100, C205S158000, C204S165000
Reexamination Certificate
active
06197949
ABSTRACT:
The present invention relates to the binding of nucleic acids to an electrically conductive polymer (ECP).
In a great many techniques commonly used in biology, for example the synthesis or hybridization of nucleic acids, oligonucleotides are covalently bound at their end to a solid support. Various supports have been used for this purpose: paper, nylon, glass, silica, polystyrene, polyacrylamide, etc.
At the present time, many teams are researching into the production of supports bearing a large number of oligonucleotides with various sequences, arranged according to a preestablished arrangement, in order simultaneously to perform various reactions (hybridization on a support for example).
Thus, this approach has been proposed, for example, in order to facilitate the sequencing of nucleic acids.
Various oligonucleotides arranged in columns and rows on microsurfaces (oligonucleotide matrices on a support) have been proposed in order to sequence nucleic acids [LYSOV et al., Proc. USSR Acad. Sci., 303, 1508-1511, (1988); KHRAPKO et al., FEBS Lett. 256, 118-122, (1989); KHRAPKO et al., DNA Séquence, vol 1, 375-388 (1991); BAINS & SMITH, J.Theor.Biol. 135, 303-307, (1988); CHURCH & KIEFFER-HIGGINS, Science 240, 185-188 (1988); SOUTHERN, PCT application WO89/10977 (1989)]. The method is based on the hybridization of target DNA or RNA chains on a set of oligonucleotides. In theory, the presence or absence of a sequence in the target nucleic acid may be determined by the hybridization observed on the microsurfaces under defined rigorous conditions.
As regards the in situ synthesis of polynucleotides or polypeptides, by combining the methods of chemical synthesis on a solid phase, photolabile groups and photolithography, FODOR et al. [Science, 251, 767-773 (1991)] have succeeded in synthesizing 1024 peptides on a grid (square, side length 100 &mgr;m). These peptides were obtained by simultaneous and parallel syntheses, using photolithography masks and photolabile protecting groups for the peptide synthons. A dCpT dinucleotide was prepared in situ, using thymidine which was 5′-protected by a photolabile protecting group (5′-nitroveratryl thymidine). The light was directed by a photolithography mask and a deposit in a checked pattern with a side length of 100 &mgr;m was obtained.
MASKOS & SOUTHERN (Nucleic Acids Res. 1992, 20, 1675-1678) performed, under a microscope, the in situ synthesis of four different oligonucleotides on a glass slide.
Hitherto, the techniques used for the directed deposition of oligonucleotides use either manual deposition (which cannot be used on the industrial scale), or photolithography techniques, which require the use of “masks” and are, moreover, difficult to apply with nucleic acids, which are photolabile.
The aim of the present invention is to obtain novel supports and novel processes for binding oligonucleotides, which do not have the drawbacks of the processes proposed in the prior art.
With this aim, the inventors have had the idea of using electrically conductive polymers as a binding support.
The inventors are now able to bind stably, and via a covalent bond, nucleotides and oligonucleotides to an electrically conductive polymer, and thereby to obtain novel copolymers.
The subject of the present invention is a copolymer which corresponds to the following general formula (I):
in which the unit A represents a monomer of an electrically conductive polymer, the unit B represents a nucleotide, an oligonucleotide or one of the analogues thereof, x, y and z represent integers equal to or greater than 1, or y may be equal to 0, and l represents a covalent bond or a spacer arm.
REFERENCES:
patent: 5837859 (1998-11-01), Teoule et al.
patent: 0314009A3 (1989-05-01), None
patent: WO 91/08307 (1991-06-01), None
Barthet Christelle
Bidan Gerard
Livache Thierry
Roget Andre
Teoule Robert
Cis Bio International
Kafafi Sherif
Marschel Ardin H.
Oblon & Spivak, McClelland, Maier & Neustadt P.C.
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