Modification of polymer surfaces and molecular immobilization by

Details Modification of polymer surfaces and molecular immobilization by Modification of polymer surfaces and molecular immobilization by

1990-07-10

1995-06-27

Webman, Edward J.

Drug, bio-affecting and body treating compositions

Solid synthetic organic polymer as designated organic active...

Aftertreated polymer

424 7819, 424 7824, 424490, 424491, 424493, 427508, 427 21, 522 84, 522 93, 522 95, 530815, 530816, A61K 4100, A61K 4732, A61K 4734, A61K 916

Patent

active

054277795

DESCRIPTION:

BRIEF SUMMARY
BRIEF DESCRIPTION OF THE INVENTION

The present invention relates to a method for modifying the surface of a polymer. In particular, the invention relates to a method for immobilizing compounds on a solid polymer phase, preferably polystyrene, polyvinyl chloride or polyethylene terephthalate glycol.


BACKGROUND OF THE INVENTION

Immobilization of biomolecules on solid phases is widely used in numerous techniques such as e.g. in chromatography, in bio-sensors, in bio-reactors, e.g. for solid phase enzyme processing, chemical synthesis of peptides, oligonucleotides or other compounds and in so-called heterogeneous immunoassays.
In the heterogeneous immunoassays, antigens or antibodies can be covalently coupled to carriers such as cellulose, agarose or polyacrylamide. However, when the compounds are to be bound on a solid phase, usually polystyrene, polypropylene or polyvinylchloride test tubes or micro-titre-plates, physical adsorption of the compounds has been the normal coupling method in heteregeneous immunoassays (Engvall and Pearlmann, J. Immunol., 1972, p. 109-129).
Passive physical adsorption is, however, not irreversible (Lehtonen et al., J. Immunol. Methods, 1980, p. 34-61), which may affect the reproducibility of the immunoassays, especially when the antigen/antibody coated solid phase is stored in dry form. Also, solid phase adsorbed antigen may not be recognized by its corresponding antibody because of denaturation (reconformation) of the antigen tertiary structure (Kurki and Virtanen, J. Immunol. Methods, 1984, p. 67-122). The antigen may also exhibit new antigenic determinants when denatured, as in the case of DNA.
The ability of passive binding to plastic is furthermore restricted to a limited amount of molecules such as e.g. proteins, or single-stranded DNA. However, some proteins and nucleic acids as well as polysaccharides and smaller molecules cannot be adsorbed directly to some types of plastic.
Covalent binding, in contrast to simple physical binding, orientates all immobilized compounds in a defined way on the solid phase, thereby exposing defined areas e.g. antigens, antibodies or enzyme catalytic sites to the fluid phase eventually poured on the solid phase surface. Antigen epitopes or active sites on these compounds can in this way remain functional. Irreversible immobilization of molecules may furthermore have some advantages in relation to storage of solid phase immobilized compounds in their dry state, since passive physical binding partly denatures some proteins and nucleic acids.
For all these reasons it would be advantageous if it was possible to immobilize antigens, antibodies or other molecules covalently to the solid phase.
Covalent solid phase fixation of some types of compounds to polystyrene has been known for several years. Solid phase peptide and oligonucleotide syntheses are performed, for example, using modified polystyrene particles as the solid support. However, these modification methods frequently involve very hazardous chemicals and several time-consuming operation steps. An example of this is the preparation of Merrifield's peptide resin which has been widely used in peptide synthesis. Preparation of this resin involves the extremely carcinogenic reagent chloromethylmethyl ether and this and other organic solvents often result in a turbid surface because the plastic is slightly soluble in the reagents. This is inconvenient if a subsequent spectrophotometrical detection is desired.
Polystyrenes premodified with, for example, --OH, --SO.sub.3 H or --NH.sub.2 groups are available. When using these premodified polystyrenes a separate production of particles for each type of modified polystyrene is necessary.
A method for introducing amino groups on plastic surfaces for use in micro-titre-plates has been described (J. Virol. Methods 3, 1981, p. 155-165). This procedure, which requires the hazardous chemicals methanesulphonic acid, glacial acetic acid and fuming nitric acid, takes two days and needs a well functioning hood. Such conditions can hardly be used in large

REFERENCES:
patent: 3625745 (1971-12-01), Wright

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