Biochemistry reactive material and device for eliminating electronegative low-density lipoprotein (ldl) and method for treating blood or plasma ex vivo to eliminate electronegative low-density lipoprotein therein

Abstract

The present disclosure provides a biochemistry reactive material, including a substrate and an enzyme composition immobilized on the substrate. The enzyme composition is selected from a group consisting of a first enzyme, a second enzyme, and a combination thereof. The first enzyme is used for eliminating a glycan residue of an electronegative low-density lipoprotein (electronegative LDL). The second enzyme is used for eliminating ceramide carried by an electronegative low-density lipoprotein. The biochemistry reactive material is capable of eliminating electronegative low-density lipoprotein.

What is claimed is: 
     
         1 . A biochemistry reactive material, comprising:
 a substrate; and   an enzyme composition immobilized on the substrate, wherein the enzyme composition is selected from a group consisting of:
 a first enzyme for eliminating a glycan residue of an electronegative low-density lipoprotein (electronegative LDL); 
 a second enzyme for eliminating ceramide carried by an electronegative low-density lipoprotein; and 
 a combination thereof, 
   wherein the biochemistry reactive material is capable of eliminating electronegative low-density lipoprotein.   
     
     
         2 . The biochemistry reactive material as claimed in  claim 1 , wherein the substrate comprises silica gel, cellulose, diethylaminoethyl cellulose (DEAE cellulose), chitosan, polystyrene, polysulfone, polyethersulfone, acrylate resin or polysaccharide. 
     
     
         3 . The biochemistry reactive material as claimed in  claim 1 , wherein the substrate has a particle structure or a hollow-tube structure. 
     
     
         4 . The biochemistry reactive material as claimed in  claim 1 , wherein the substrate is a cellulose bead. 
     
     
         5 . The biochemistry reactive material as claimed in  claim 1 , wherein the substrate is a chitosan bead. 
     
     
         6 . The biochemistry reactive material as claimed in  claim 1 , wherein the substrate is a cellulose hollow fiber, a polysulfone hollow fiber, epoxy acrylic resin or a polyethersulfone hollow fiber. 
     
     
         7 . The biochemistry reactive material as claimed in  claim 1 , wherein the first enzyme is sialidase or glycosidase. 
     
     
         8 . The biochemistry reactive material as claimed in  claim 7 , wherein the sialidase is selected from a group consisting of:
 neuraminidase 1 (NEU1), neuraminidase 2 (NEU2), neuraminidase 3 (NEU3), neuraminidase 4 (NEU4) and O-sialidase bioengineered from human genome, one of the foregoing enzymes obtained through gene transformation, expression and purification, and sialidase from a virus or bacterium (alias, acetylneuraminyl hydrolase).   
     
     
         9 . The biochemistry reactive material as claimed in  claim 7 , wherein the glycosidase is selected from a group consisting of:
 alpha- and beta-glucosidase bioengineered from human or animal genome, maltase-glucoamylase and sucrase-isomaltase, one of the foregoing enzymes obtained through gene transformation, expression and purification, and N-glycosidase F (PNGase F) and glucosidase from a virus or bacterium.   
     
     
         10 . The biochemistry reactive material as claimed in  claim 1 , wherein the second enzyme is ceramidase. 
     
     
         11 . The biochemistry reactive material as claimed in  claim 10 , wherein the ceramidase is selected from a group consisting of:
 N-acylsphingosine amidohydrolase 1 (ASAH1), N-acylsphingosine amidohydrolase 2 (ASAH2), N-acylsphingosine amidohydrolase 2B (ASAH2B), N-acylsphingosine amidohydrolase 2C (ASAH2C), N-acylethanolamine acid amidase, alkaline ceramidase 1, alkaline ceramidase 2 and alkaline ceramidase 3.   
     
     
         12 . The biochemistry reactive material as claimed in  claim 1 , wherein the enzyme composition is the first enzyme. 
     
     
         13 . The biochemistry reactive material as claimed in  claim 12 , wherein the first enzyme is neuraminidase 2. 
     
     
         14 . The biochemistry reactive material as claimed in  claim 1 , wherein the enzyme composition is the second enzyme. 
     
     
         15 . The biochemistry reactive material as claimed in  claim 14 , wherein the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         16 . The biochemistry reactive material as claimed in  claim 1 , wherein the enzyme composition is the combination of the first enzyme and the second enzyme. 
     
     
         17 . The biochemistry reactive material as claimed in  claim 16 , wherein the first enzyme is neuraminidase 2, and the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         18 . The biochemistry reactive material as claimed in  claim 1 , wherein the electronegative low-density lipoprotein comprises electronegative low-density lipoprotein L1, L2, L3, L4 or L5. 
     
     
         19 . The biochemistry reactive material as claimed in  claim 18 , wherein the electronegative low-density lipoprotein is electronegative low-density lipoprotein L5. 
     
     
         20 . A biochemistry reactive device, comprising:
 the biochemistry reactive material as claimed in  claim 1 ; and   a container for containing the biochemistry reactive material, wherein the container has at least one inlet and at least one outlet,   wherein a liquid sample enters into the biochemistry reactive device from the at least one inlet, and flows through the biochemistry reactive material to react with the biochemistry reactive material, and then flows out through the at least one outlet.   
     
     
         21 . The biochemistry reactive device as claimed in  claim 20 , wherein a material of the container comprises glass, acrylic, polypropylene, polyethylene, stainless steel or titanium alloy. 
     
     
         22 . The biochemistry reactive device as claimed in  claim 20 , further comprising:
 a filtering material configured in the container behind the at least one inlet and at least one outlet,   wherein a pore size of the filtering material is smaller than the biochemistry reactive material to prevent the biochemistry reactive material leaking from the at least one inlet and/or least one outlet.   
     
     
         23 . The biochemistry reactive device as claimed in  claim 20 , wherein a material of the filtering material comprises filter paper, glass, acrylic, polypropylene or polyethylene. 
     
     
         24 . The biochemistry reactive device as claimed in  claim 20 , wherein the container is a hollow column, and two ends of the container have a first inlet of the at least one inlet and a first outlet of the at least outlet, respectively. 
     
     
         25 . The biochemistry reactive device as claimed in  claim 24 , a second inlet of the at least one inlet and a second outlet of the at least outlet are located at a side wall of the hollow column. 
     
     
         26 . The biochemistry reactive device as claimed in  claim 20 , wherein the substrate has a particle structure or a hollow-tube structure. 
     
     
         27 . The biochemistry reactive device as claimed in  claim 22 , wherein the substrate has a particle structure or a hollow-tube structure. 
     
     
         28 . The biochemistry reactive device as claimed in  claim 22 , wherein the substrate has a particle structure. 
     
     
         29 . The biochemistry reactive device as claimed in  claim 24 , wherein the substrate has a particle structure or a hollow-tube structure. 
     
     
         30 . The biochemistry reactive device as claimed in  claim 25 , wherein the substrate has a hollow-tube structure. 
     
     
         31 . The biochemistry reactive device as claimed in  claim 20 , wherein the substrate comprises silica gel, cellulose, diethylaminoethyl cellulose, chitosan, polystyrene, polysulfone, polyethersulfone, acrylate resin or polysaccharide. 
     
     
         32 . The biochemistry reactive device as claimed in  claim 28 , wherein the substrate is a cellulose bead. 
     
     
         33 . The biochemistry reactive device as claimed in  claim 28 , wherein the substrate is a chitosan bead. 
     
     
         34 . The biochemistry reactive device as claimed in  claim 30 , wherein the substrate is a cellulose hollow fiber, a polysulfone hollow fiber, epoxy acrylic resin or a polyethersulfone hollow fiber. 
     
     
         35 . The biochemistry reactive device as claimed in  claim 20 , wherein the first enzyme is sialidase or glycosidase. 
     
     
         36 . The biochemistry reactive device as claimed in  claim 35 , wherein the sialidase is selected from a group consisting of:
 neuraminidase 1 (NEU1), neuraminidase 2 (NEU2), neuraminidase 3 (NEU3), neuraminidase 4 (NEU4) and O-sialidase bioengineered from human genome, one of the foregoing enzymes obtained through gene transformation, expression and purification, and sialidase from a virus or bacterium (alias, acetylneuraminyl hydrolase).   
     
     
         37 . The biochemistry reactive device as claimed in  claim 35 , wherein the glycosidase is selected from a group consisting of:
 alpha- and beta-glucosidase bioengineered from human or animal genome, maltase-glucoamylase and sucrase-isomaltase, one of the foregoing enzymes obtained through gene transformation, expression and purification, and N-glycosidase F (PNGase F) and glucosidase from a virus or bacterium.   
     
     
         38 . The biochemistry reactive device as claimed in  claim 20 , wherein the second enzyme is ceramidase. 
     
     
         39 . The biochemistry reactive device as claimed in  claim 38 , wherein the ceramidase is selected from a group consisting of:
 N-acylsphingosine amidohydrolase 1, N-acylsphingosine amidohydrolase 2, N-acylsphingosine amidohydrolase 2B, N-acylsphingosine amidohydrolase 2C, N-acylethanolamine acid amidase, alkaline ceramidase 1, alkaline ceramidase 2 and alkaline ceramidase 3.   
     
     
         40 . The biochemistry reactive device as claimed in  claim 20 , wherein the enzyme composition is the first enzyme, and the first enzyme is neuraminidase 2. 
     
     
         41 . The biochemistry reactive device as claimed in  claim 20 , wherein the enzyme composition is the second enzyme, and the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         42 . The biochemistry reactive device as claimed in  claim 20 , wherein the enzyme composition is the combination of the first enzyme and the second enzyme, and the first enzyme is neuraminidase 2 and the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         43 . The biochemistry reactive device as claimed in  claim 20 , wherein the electronegative low-density lipoprotein comprises electronegative low-density lipoprotein L1, L2, L3, L4 or L5. 
     
     
         44 . The biochemistry reactive device as claimed in  claim 20 , wherein the electronegative low-density lipoprotein is electronegative low-density lipoprotein L5. 
     
     
         45 . The biochemistry reactive device as claimed in  claim 20 , wherein the liquid sample comprises aqueous solution, blood or plasma. 
     
     
         46 . A method for ex vivo treating blood or plasma, comprising:
 (a) ex vivo contacting a blood or plasma with an enzyme composition to react the enzyme composition with the blood or plasma, wherein the enzyme composition is capable of eliminating electronegative low-density lipoprotein, and the enzyme composition is selected from a group consisting of:
 a first enzyme for eliminating a glycan residue of an electronegative low-density lipoprotein (LDL); 
 a second enzyme for eliminating ceramide carried by a electronegative low-density lipoprotein (LDL); and 
 a combination thereof; and 
   (b) terminating contact between the blood or plasma and the enzyme composition to terminate the reaction of the enzyme composition with the blood or plasma.   
     
     
         47 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the step (a) is performed for about 0.25-8 hours. 
     
     
         48 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the step (a) is performed at about 4-40° C. 
     
     
         49 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the step (a) is performed at about pH 5-10. 
     
     
         50 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the first enzyme is sialidase or glycosidase. 
     
     
         51 . The method for ex vivo treating blood or plasma as claimed in  claim 50 , wherein the sialidase is selected from a group consisting of:
 neuraminidase 1 (NEU1), neuraminidase 2 (NEU2), neuraminidase 3 (NEU3), neuraminidase 4 (NEU4) and O-sialidase bioengineered from human genome, one of the foregoing enzymes obtained through gene transformation, expression and purification, and sialidase from a virus or bacterium (alias, acetylneuraminyl hydrolase).   
     
     
         52 . The method for ex vivo treating blood or plasma as claimed in  claim 50 , wherein the glycosidase is selected from a group consisting of:
 alpha- and beta-glucosidase bioengineered from human or animal genome, maltase-glucoamylase and sucrase-isomaltase, one of the foregoing enzymes obtained through gene transformation, expression and purification, and N-glycosidase F (PNGase F) and glucosidase from a virus or bacterium.   
     
     
         53 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the second enzyme is ceramidase. 
     
     
         54 . The method for ex vivo treating blood or plasma as claimed in  claim 53 , wherein the ceramidase is selected from a group consisting of:
 N-acylsphingosine amidohydrolase 1, N-acylsphingosine amidohydrolase 2, N-acylsphingosine amidohydrolase 2B, N-acylsphingosine amidohydrolase 2C, N-acylethanolamine acid amidase, alkaline ceramidase 1, alkaline ceramidase 2 and alkaline ceramidase 3.   
     
     
         55 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the enzyme composition is the first enzyme. 
     
     
         56 . The method for ex vivo treating blood or plasma as claimed in  claim 55 , wherein the first enzyme is neuraminidase 2. 
     
     
         57 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the enzyme composition is the second enzyme. 
     
     
         58 . The method for ex vivo treating blood or plasma as claimed in  claim 57 , wherein the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         59 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the enzyme composition is the combination of the first enzyme and the second enzyme. 
     
     
         60 . The method for ex vivo treating blood or plasma as claimed in  claim 59 , wherein the first enzyme is neuraminidase 2, and the second enzyme is N-acylsphingosine amidohydrolase 2. 
     
     
         61 . The method for ex vivo treating blood or plasma as claimed in  claim 46 , wherein the enzyme composition is immobilized on the substrate. 
     
     
         62 . The method for ex vivo treating blood or plasma as claimed in  claim 61 , the substrate comprises silica gel, cellulose, diethylaminoethyl cellulose, chitosan, polystyrene, polysulfone, polyethersulfone, resin or polysaccharide. 
     
     
         63 . The method for ex vivo treating blood or plasma as claimed in  claim 61 , the substrate has a particle structure or a hollow-tube structure.