Biomimetic particles and films for pathogen capture and other uses

Abstract

The identification and immobilization of glycosylated molecules having biomimetic properties, more particularly naturally-occurring, tissue-derived, non-immunological glycan sequences or functional equivalents thereof, on solid state surfaces and films or on membranes arising at the interface between non-polar and polar materials is described herein. The biomimetic glycosylated films and particles constructed therefrom have industrial, environmental, diagnostic and/or therapeutic utility in the binding, capture, and/or extraction of pathogens, toxins and/or contaminants, in vivo, in vitro or in situ. The present invention further extends to the use of such biomimetic films and particles for the delivery of other therapeutic molecules as well as in the construction of body contacting devices having enhanced biocompatibility and reduced immunogenicity.

1 - 97 . (canceled) 
     
     
         98 . A solid state film with biomimetic properties having at least one functionally active surface, said film comprising a substrate having a plurality of glycosylated molecules immobilized therein, wherein said glycosylated molecules are either harvested or derived from biological fluid or tissue, or comprise a functional mimic of a component of said biological fluid or tissue. 
     
     
         99 . The biomimetic film of  claim 98 , wherein the glycosylated molecules comprise endogenous cell-surface glycan signatures harvested or derived from biological fluid or tissue or functional mimics thereof. 
     
     
         100 . The biomimetic film of  claim 99 , wherein said glycan signature are selected from the group consisting of pathogen binding moieties and self antigens that masks the foreign nature of the film and thereby reducing its immunogenic potential. 
     
     
         101 . The biomimetic film of  claim 98 , wherein the glycosylated molecules are selected from the group consisting of glycoproteins, glycolipids, glycosides, glycosylated transmembrane proteins, glycodendrimers, glycodendriproteins, and non-immunological sialated glycosylated molecules. 
     
     
         102 . The biomimetic film of  claim 98 , wherein the glycosylated molecules include surface moieties of attaching and effacing (A/E) lesions or functional mimics thereof. 
     
     
         103 . The biomimetic film of  claim 99 , wherein said glycan signatures are harvested or derived from the inner or outer walls or membranes of the organs of the gastrointestinal tract, urinary tract, pulmonary tract, blood vessels, amniotic sac, ocular sac, nervous system, muscoskeletal system. 
     
     
         104 . The biomimetic film of  claim 99 , wherein said glycan signatures are isolated from body fluids selected from the group consisting of mucous, blood, blood plasma, saliva, urine, synovial fluid, breast milk, tears, fluids of the reproductive system, aqueous or vitreous humor, bone marrow, and cerebrospinal fluids. 
     
     
         105 . The biomimetic film of  claim 98 , wherein said substrate is non-polar or exhibits hydrophobic surface properties and is selected from the group consisting of resin, natural wax, synthetic wax, natural and synthetic plastics, polymers, hydrocarbon mixtures, natural and synthetic fats, ceramics, glass, diatomaceous earth, and metal. 
     
     
         106 . The biomimetic film of  claim 98 , wherein said substrate further comprises a detectable label selected from the group consisting of a fluorescent label, a radioactive label, a dye, and a compound that enhances magnetic resonance imaging. 
     
     
         107 . The solid state biomimetic film of  claim 98 , wherein said film comprises a relatively non-polar substrate and said plurality of glycosylated molecules are immobilized to the upper surface of said substrate, further wherein one or more amphiphilic molecules having hydrophobic tail ends and hydrophilic head ends are embedded in or chemically, electrically or mechanically linked via said hydrophobic tail ends to the relatively non-polar substrate, further wherein the amphiphilic molecules either themselves comprise biomimetic moieties or have such biomimetic moieties attached or conjugated to their hydrophilic head ends such that the biomimetic moieties project from said upper surface so as to provide at least one surface of said film with functional activity. 
     
     
         108 . The solid state biomimetic film of  claim 107 , wherein a heterogeneous mixture of amphiphilic molecules or conjugates with pathogen and/or toxin binding properties is present in the film in a manner which provides for multiple pathogen and/or toxin binding mechanisms or moieties. 
     
     
         109 . A therapeutic biocompatible particle formed from the solid state biomimetic film of claim  1 , said particle of a size or geometric configuration that prevents its absorption by the biological tissue of an organism. 
     
     
         110 . The therapeutic particle of  claim 109 , wherein said biological tissue is selected from the group consisting of biological tissue extracted from human, animal or plant tissue; a culture of human, animal or plant tissue; a biological surrogate for human, animal or plant tissue; a culture of a biological surrogate for human, animal or plant tissue or a recombinant version of human, animal or plant tissue or a human, animal or plant tissue biological surrogate, further wherein said biological tissues are selected from the group consisting of epithelium, connective tissue, muscle tissue, nerve tissue, material associated with or contained in amniotic fluid surrounding a fetus, aqueous humour, blood, blood plasma, interstitial fluid, breast milk, mucus, pus, saliva, serum, tears, urine, cerebrospinal fluid, synovial fluid, intracellular fluid, aqueous humour, vitreous humour and other bodily fluids. 
     
     
         111 . The therapeutic particle of  claim 109 , wherein the substrate comprises a nonbiodegradable material capable of maintaining the immobilization of said glycosylated molecules when ingested by a living organism. 
     
     
         112 . The therapeutic particle of  claim 109 , wherein the particle is in the form of a wax micelle. 
     
     
         113 . The therapeutic particle of  claim 109 , wherein the particle is in the form of a worm micelle. 
     
     
         114 . A pharmaceutical composition comprising the therapeutic particle of  claim 109  formulated for introduction into a living organism. 
     
     
         115 . The pharmaceutical composition of  claim 114 , wherein said composition further comprises one or more additional agents selected from the group consisting of antibiotic agents, probiotics, vitamin supplements such as iron, or therapeutic agents for oral rehydration therapy such as oral rehydration salts, solution, and electrolytes. 
     
     
         116 . A method for making the biomimetic film of  claim 98  comprising the following steps, with the resultant films being disposed on solid substrates:
 a. providing a relatively polar solvent having sufficient quantities dissolved therein of one or more amphiphilic molecules which include glycosylated molecules with biomimetic properties, or conjugates of such, or molecules which can be c conjugated with glycosylated molecules with biomimetic properties, or molecules with other pathogen binding moieties;   b. exposing the polar solvent to a relatively non-polar liquid, the non-polar liquid being immiscible in the polar solvent; the relatively non-polar liquid can optionally comprise a material which can reversibly experience a state change from solid to a relatively non-polar liquid under stimulus;   c. allowing the amphiphilic molecules to align so as to form a membrane which separates the polar solvent from the non-polar liquid; and   d. inducing or allowing transformation of the non-polar liquid to a corresponding non-polar solid having upper and lower surfaces, wherein the hydrophobic tail ends of the amphiphilic molecule are embedded in or chemically, electrically or mechanically linked to the upper surface of the non-polar substrate and the hydrophilic tail ends project from the upper surface into the polar solvent so as to yield a film having functional activity.   
     
     
         117 . The method of  claim 116 , further comprising the step of adsorbing or conjugating molecules with biomimetic or other properties to the functional surface of the film if the desired functionality is not present from the preceding steps. 
     
     
         118 . A method of forming the biomimetic film of  claim 98  from biological cell- or membrane-bound glycosylated molecules comprising the following steps:
 a. isolating biological tissue extracted from human, animal or plant tissue, a culture of human, animal or plant tissue, a biological surrogate for human, animal or plant tissue, a culture of a biological surrogate for human, animal or plant tissue or recombinant versions of human, animal or plant tissue or human, animal or plant tissue biological surrogates;   b. subjecting the biological cell or membrane to a process which ruptures or lyses the cell or other membrane, including but not limited to, mechanical means (for example, but not limited to, sonication, freeze/thaw and high shear techniques), chemical means (for example, but not limited to, using detergents for whole cell lysis and cell fractionation) and biochemical means (for example, but not limited to, by means of enzymes and/or protease inhibitors), to form a liquid volume with biomimetic molecules present;   c. optionally filtering or otherwise selectively separating a fraction of interest containing glycosylated molecules with biomimetic properties, said fraction may or may not comprise a liquid or other molecules;   d. immobilizing said biomimetic molecules as part of a film on an appropriate substrate; and   e. optionally removing said film from any surrounding liquid to form a dry construct.   
     
     
         119 . The method of  claim 118 , wherein the step of selectively separating a fraction of interest containing glycosylated molecules with biomimetic properties comprises the following steps for molecules which are amphiphilic in nature:
 a. exposing the solution containing biological tissue, which can comprise, in part, lysed or otherwise ruptured cell material containing molecules of interest in the present invention to a less-polar material;   b. if necessary, adding amphiphilic molecules to achieve critical micelle concentration, before or after exposing said solution to said less-polar material;   c. allowing the amphiphilic compounds to form a film at the interface between said solution and said less-polar material;   d. conducting any desired procedures to affix the molecules to the less-polar substrate beyond the hydrophobic forces inherent in the film; and   e. removing, or replacing with another liquid, some or all of said solution.   
     
     
         120 . The method of  claim 118 , wherein the transformation is achieved through cooling of the non-polar liquid. 
     
     
         121 . A method of using of biomimetic film of  claim 98  comprising the steps of:
 a. exposing said surface to a liquid sample which may or may not contain pathogens or toxins;   b. allowing sufficient time for said surface to bind to target pathogens or toxins present in said sample;   c. optionally separating said surface, to which said target pathogens or toxins may be bound, from said sample; and   d. optionally, analyzing said surface to detect the presence or type of pathogen or toxin, or subjecting said surface, to which said target pathogens or toxins may be bound, to an agent which effectively separates the pathogens or toxins from the surface and subjecting said pathogen or toxins, or media containing such, to analysis techniques to detect presence or type.   
     
     
         122 . A method of using the biomimetic film of  claim 98 , comprising the steps of:
 a. extracting of a bodily fluid of an organism, including, but not limited to amniotic fluid, aqueous or vitreous humour, blood and blood plasma, bone marrow fluids, cerebrospinal fluid, interstitial fluid, lymph fluids and pleural fluid;   b. inducing contact of said bodily fluid to one or more surfaces with the pathogen and toxin binding moieties of the present invention;   c. optionally, reintroducing of the bodily fluid into the organism; and   d. optionally, subjecting said surfaces of the present invention to analysis techniques to determine presence and/or type of pathogens or toxins that may have bound to the surface.   
     
     
         123 . A method of using the therapeutic particle of  claim 109 , comprising the steps of:
 a. administering one or multiple oral doses of a plurality of particles of constructs of the present invention to the host;   b. allowing the particles of the present invention to contact areas of the alimentary system of the host where pathogens may be present; and   c. allowing the host organism to discharge the particles through defecation, vomiting, expectoration or other means of natural or induced discharge.   
     
     
         124 . A method of using the therapeutic particle of  claim 109 , comprising the steps of:
 a. administering one or multiple doses of a plurality of particles of the present invention to the host through a catheter or similar device or through other means of physically placement into a region of suspected infection or a point in the host where the particles will migrate to a region where pathogens may be present;   b. allowing the particles of the present invention to contact those areas of the host where pathogens may be present; and   c. allowing the host organism to discharge the particles through defecation, urination, vomiting, mucosal flow, expectoration or other means of natural or induced discharge or removing the particles from the host through a catheter or other device.   
     
     
         125 . A medical device with enhanced surface biocompatibility formed from or coated with the biomimetic film of  claim 98 . 
     
     
         126 . The device of  claim 125 , wherein said biomimetic glycosylated molecules are harvested or derived from tissue of an intended host organism of said medical device or an antigen-matched donor thereof or from a culture of tissue from said recipient or donor.