Carleton University

Other

Burton W Blais

Hiroshi Yamazaki

An improved device is provided which may be used, e.g. for immunoassay of lipopolysaccharides or for removing LPS pyrogens from aqueous solutions, or for removing LPS endotoxins from wounds. Such device comprises, in combination, a substrate, e.g. plastic, i.e. polystyrene, polycarbonate, polymethylmethacrylate or polyvinyl chloride, or a woven cloth, i.e. a rayon/polyester cloth or a polyester cloth, or a non-woven cloth, i.e. a rayon/polyester cloth, or a polyester cloth, or paper, which is adapted to receive a sample to be tested, and an oligopeptide, or a hydrophobic polypeptide or a polymyxin, e.g. polymyxin B, polymyxin B.sub.1, polymyxin B.sub.2, polymyxin D.sub.1, polymyxin D.sub.2, or polymyxin E, adhered to the substrate.

Method for using polymyxin-coated substrate for lipopolysaccharide

A device for immunological techniques is prepared containing a macroporous hydrophobic synthetic polymer cloth having antibodies or antigens directly adsorbed therein and directly absorbed and immobilized thereon. The cloth has a thickness of more than about 200 .mu.m and has spaces between fibres exceeding about 20 .mu.m in diameter, and preferably has a Frazier Air permeability, in CFM/ft.sup.2 at 0.5' H.sub.2 O of from about 215 to about 750 for thickness of from about 11 to about 40 mils such that it can accommodate a large volume of liquid per surface area, that it has a large surface area, and that it has minimum flow resistance. In immunoassays antibodies may be directly adsorbed therein and directly absorbed and immobilized thereon, and specific antigens from a selected test sample, may then be captured by the antibodies, to be detected conventionally. Alternatively antigens may be directly adsorbed therein and directly absorbed and immobilized thereon, and specific antibodies from a selected test sample, may be captured by the antigens, to be detected conventionally. This device can also be used for extraction and concentration of lipopolysaccharide antigens and for preparing biotinylated, affinity-purified antibodies.

Antibodies or antigens bound to a macroporous hydrophobic synthetic

An immunoassay device containing an immobilized antibody or antigen is provided by directly absorbing and absorbing an unmodified antibody or antigen on and within a woven or non-woven macroporous hydrophobic synthetic polymer cloth formed of a synthetic polymer selected from the group consisting of plypropylene, polyester, nylon and polyethylene. The cloth has a thickness of more than about 200 .mu.m and contains pores in the form of spaces between fibers exceeding about 20 .mu.m in diameter, and has a Frazier Air Permeability in CFM/ft.sup.2 at 0.5' H.sub.2 O of about 215 for a cloth of thickness of about 40 mils. The cloth has a large surface area for binding to an antibody or antigen and can accommodate a large volume of liquid per surface area and has minimum flow resistance. The cloth containing an immobilized antibody or antigen may be used to carry out an enzyme immunoassay by contacting the cloth with a sample containing an antigen or antibody, incubating the cloth with an enzyme-antibody conjugate and then reacting enzyme bound to the cloth with a chromogenic substrate-indicator to produce a visible color. Other immunoassay embodiments may also be carried out and a control cloth can be used such that a difference in color from that obtained with the control cloth determines the amount of antigen or antibody present in a sample.

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Ottawa, Canada

Burton W Blais