Method for biomaterial purification and kits thereof
Described herein is a method of purifying a target molecule from an aqueous biological composition, the method comprising: (a) contacting a cationic polymer and the aqueous biological composition to form a mixture comprising a bio-polymer complex and the target molecule in a liquid, wherein the bio-polymer complex has an average particle diameter of at least 45 micrometers, (b) adding the mixture to a filtering volume of a vessel, wherein the vessel comprises loosely packed staple fibers; (c) allowing the mixture to separate through the loosely packed staple fiber; and (d) collecting a filtrate comprising the target molecule.
1 . A method of purifying a target non-binding molecule from an aqueous biological composition comprising a binding species, the method comprising:
(a) contacting a cationic polymer and the aqueous biological composition to form a mixture comprising a bio-polymer complex and the target non-binding molecule in a liquid, wherein the bio-polymer complex has an average particle diameter of at least 45 micrometers,
(b) adding the mixture to a filtering volume of a vessel, wherein the vessel comprises loosely packed staple fibers;
(c) allowing the mixture to separate through the loosely packed staple fibers; and
(d) collecting a filtrate comprising the target non-binding molecule;
wherein said loosely packed staple fibers are not cationically functionalized.
2 . The method of claim 1 , wherein the loosely packed staple fibers have a packing density of at least 0.03 g/cm 3 .
3 . The method of claim 1 , wherein the loosely packed staple fibers have a packing density of at most 0.24 g/cm 3 .
4 . The method of claim 1 , wherein the cationic polymer is a water soluble or water dispersible polymer.
5 . The method of claim 1 , wherein the cationic polymer is functionalized with guanidinyl groups.
6 . The method of claim 5 , wherein the cationic polymer comprises groups of the formula:
—[C(R 1 )═N—R 2 ] n —N(R 3 )—[C(═N—R 4 )N(R 4 )] m —R 5
wherein
R 1 is a H, C1-C12 alkyl, C5-C12 (hetero)aryl, or a residue of the polymer chain;
R 2 is a covalent bond, a C2-C12 (hetero)alkylene, or a C5-C12 (hetero)arylene;
R 3 is H, C1-C12 alkyl, or C5-C12 (hetero)aryl; and
each R 4 is independently H, C1-C12 alkyl or alkylene, C5-C12 (hetero)aryl or (hetero) arylene, cyano, or —C(═NH)—N(R 2 )-Polymer;
R 5 is H, C1-C12 (hetero)alkyl, C5-C12 (hetero)alkyl, or —N(R 4 ) 2 ;
n is 0 or 1; and
m is 1 or 2.
7 . The method of claim 1 , wherein the cationic polymer is further functionalized with quaternary ammonium groups.
8 . The method of claim 1 , wherein the cationic polymer is derived from an amino polymer, optionally, wherein the amino polymer is selected from the group consisting of polyethylenimine, polylysine, polyaminoamides, polyallylamine, polyvinylamine, polydimethylamine-epichlorohydrin-ethylenediamine, and dendrimers formed from polyamidoamine (PAMAM) and polypropylenimine.
9 . The method of claim 8 , wherein 0.1 to 100 mole percent of the available amino groups of the amino polymer are functionalized with guanidinyl groups, optionally, wherein the guanidinyl groups are in the amino polymer chain.
10 . The method of claim 1 , wherein the cationic polymer is derived from a carbonyl polymer, optionally, wherein the carbonyl polymer is selected from the group consisting of; acrolein, vinyl methyl ketone, vinyl ethyl ketone, vinyl isobutyl ketone, diacetone (meth)acrylamide, acetonyl acrylate, carbon monoxide copolymer, and diacetone (meth)acrylate (co)polymers.
11 . The method of claim 1 , wherein 0.1 to 10,000 micrograms of cationic polymer is added per mL of the aqueous biological composition.
12 . The method of claim 1 , wherein the bio-polymer complex has an average particle diameter of at most 200 micrometers.
13 . The method of claim 1 , further comprising suspending the bio-polymer complex in the liquid prior to addition to the vessel.
14 . The method of claim 1 , wherein immediately following step (a), the biopolymer complex is added to the filtering volume of the vessel.
15 . The method of claim 13 , wherein suspending the bio-polymer complex in the liquid comprises agitating the bio-polymer complex in the liquid.
16 . The method of claim 1 , wherein at least a portion of the loosely packed staple fibers are hydrophilic.
17 . The method of claim 1 , wherein at least a portion of the loosely packed staple fibers are fibrillated.
18 . A kit comprising:
(a) plurality of staple fibers wherein the staple fibers are loosely packed in a bed; and
(b) a cationic polymer;
wherein said staple fibers are not cationically functionalized.
19 . The kit of claim 18 , wherein the packing density of the fibers in the bed is at least 0.03 g/cm 3 and at most 0.24 g/cm 3 .