Molecular combing process for detecting macromolecules

Details Molecular combing process for detecting macromolecules Molecular combing process for detecting macromolecules

2001-06-13

2003-04-15

Fredman, Jeffrey (Department: 1655)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

C435S007100, C435S091200, C435S004000, C536S022100, C536S023100, C536S024300, C530S350000, C530S387100

Reexamination Certificate

active

06548255

ABSTRACT:

BACKGROUND OF THE INVENTION
The present invention relates to a method for aligning macromolecules such as polymers or macromolecules with biological activity, especially DNA, or proteins. The present invention also relates to the application of this method in processes for detecting for measuring intramolecular distance, for separating and/or for assaying a macromolecule in a sample.
Controlling the conformation of macromolecules represents a major industrial challenge, for example in the manufacture of sensors or of controlled molecular assemblies, or alternatively in problems of detection and analysis. It may be useful to have an elongated molecular conformation. By way of example, in the case where polymers are grafted on a substrate, it has been proposed to extend them by the action of an electric field, a flow or with the aid of optical tweezers. In particular, in biology, the alignment of DNA—by electrophoresis (Zimmermann and Cox Nucl. Acid Res. 22, p 492, 1994), free flow (Parra and Windle, Nature Genetics, 5, p 17, 1993 and WO 93/22463) or in a gel (Schwartz et al. Science 262, p 110, 1993 and USP 33531) or with the aid of optical tweezers (Perkins et al., Science 264 p 819, 1994 and also U.S. Pat. No. 5,079,169)—opens numerous possibilities in mapping, or in the detection of pathogens.
These methods only allow in general an imperfect alignment, or alternatively a transient alignment—that is to say that relaxation of the molecule occurs once the stress disappears. In the case of optical tweezers, the method is expensive, is limited to only one molecule at a time, and is difficult to carry out by non-qualified staff.
A special technique for aligning DNA by flow after cell lysis, followed by drying, has been proposed (I. Parra and B. Windle and WO 93/22463). The alignment obtained is very imperfect and nonhomogeneous and numerous nonaligned masses are observed.
SUMMARY OF THE INVENTION
The subject of the present invention is a novel and simple method for aligning macromolecules on the surface S of a support, characterized in that the triple line S/A/B (meniscus) resulting from the contact between a solvent A and the surface S and a medium B is caused to move on the said surface S, the said macromolecules having a part, especially an end, anchored on the surface S, the other part, especially the other end, being in solution in the solvent A.
It has been observed according to the present invention that the mere passage of a meniscus over molecules of which one part is anchored on a substrate, the remainder of the molecule existing freely in solution makes it possible to align them uniformly, perpendicularly to the moving meniscus, leaving them adsorbed on the surface behind the meniscus. This phenomenon is called “molecular combing” here.


REFERENCES:
patent: 4278651 (1981-07-01), Hales
patent: 4539061 (1985-09-01), Saqiv
patent: 4562157 (1985-12-01), Lowe et al.
patent: 4683202 (1987-07-01), Mullis
patent: 4698302 (1987-10-01), Whitehead et al.
patent: 4724207 (1988-02-01), Hou et al.
patent: 4921809 (1990-05-01), Schiff et al.
patent: 5019416 (1991-05-01), Honzawa
patent: 5098977 (1992-03-01), Frautschi et al.
patent: 5102798 (1992-04-01), Guiseppi-Elie
patent: 5178947 (1993-01-01), Charmot et al.
patent: 5215882 (1993-06-01), Bahl et al.
patent: 5232829 (1993-08-01), Longiaru et al.
patent: 5237016 (1993-08-01), Ghosh et al.
patent: 5240846 (1993-08-01), Collins et al.
patent: 5330891 (1994-07-01), Sutton
patent: 5372930 (1994-12-01), Colton et al.
patent: 5447841 (1995-09-01), Gray et al.
patent: 5610287 (1997-03-01), Nikiforov et al.
patent: 5624711 (1997-04-01), Sundberg et al.
patent: 5677126 (1997-10-01), Bensimon et al.
patent: 5840862 (1998-11-01), Bensimon et al.
patent: 5846724 (1998-12-01), Bensimon et al.
patent: 5851769 (1998-12-01), Gray et al.
patent: 6130044 (2000-10-01), Bensimon et al.
patent: 6303296 (2001-10-01), Bensimon et al.
patent: 3717210 (1988-12-01), None
patent: 0 125 995 (1984-11-01), None
patent: 0 127 438 (1984-12-01), None
patent: 0 350 407 (1990-01-01), None
patent: 0 388 940 (1990-09-01), None
patent: 0 391 674 (1990-10-01), None
patent: 0 435 785 (1990-12-01), None
patent: 0 578 148 (1994-01-01), None
patent: WO93/22463 (1993-11-01), None
patent: WO97/06278 (1997-02-01), None
Nave et al, “Visualization of the polarity of isolated Titin molecules: a single globular head on a long thin rod as the M band Anchroing domain”, J. Cell Biology, vol. 109:2177-2187 (1989).*
Houmeida et al, “Studies of the interaction between Titin and Myosin”, J. Cell Biol. 131:1471-1481 (1995).*
Tskhovrebova et al, “Direct visualization of extensibility in isolated titin molecules”, J. Mol. Biol. (1997) 265:100-106.*
Preliminary Search Report, Nov. 14, 1994, FR 94/07444.
Preliminary Search Report, Nov. 14, 1994, FR 94/01574.
Zimmerman et al., “DNA Stretching on Functionalized Gold Surfaces,”Nucleic Acids Res. 22:3 (1994), pp. 492-497.
Schwartz et al., “Ordered Restriction Maps of S.CerevisiaeChromosomes Constructed by Optical Mapping,”Science262:5130 (Oct. 1993), pp. 110-114.
Parra et al., “High Resolution Visual Mapping of Stretched DNA by fluorescent Hybridization,”Nature Genetics, vol. 5 (Sep. 1993), pp. 17-21.
Lund et al., “Assessment of Methods for Covalent Binding of Nucleic Acids to Magnetic Beads, Dynabeads™, and the Characteristics of the Bound Nucleic Acids in Hybridization Reactions,”Nucleic Acids Res. 16 (1988), pp. 10861-10880.
Bensimon et al., “Alignment and Sensitive Detection of DNA by a Moving Interface”,Science, vol. 265, pp. 2096-2098 (Sep. 1994).
Keller et al.,PNAS, vol. 86, pp. 5356-5360.
Beattie et al., “Hybridization of DNA Targets . . . ”,Molecular Biotechnology, vol. 4, pp. 213-225 (1995).
Netzer et al., “A New Approach to Construction . . . ”,J. Amer. Chem. Soc., vol. 105, No. 3, pp. 674-676 (1983).
Tskhovreboa et al., “Direct Visualization of Extensibility in Isolated Titin Molecules”,J. Mol. Biology, vol. 265, pp. 100-106 (1997).

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