Ultra-stable protein ionic liquids
A method for creating a stable protein/antibody ionic liquid, comprising: (a) cationizing aqueous proteins/antibodies by addition of an excess of a positively-charged crosslinker in the presence of a coupling reagent; (b) purifying the cationized proteins/antibodies; (c) titrating the cationized proteins/antibodies with a corresponding biologically-compatible counter anionic polymer to create at least one protein/antibody cation/anion pair in aqueous solution until the cation/anion pair solution becomes negative by zeta potential measurement; (d) repeatedly dialyzing the protein/antibody cation/anion pair in water to remove excess anionic polymer using at least one molecular weight cutoff 7000 dialysis membrane; (e) lyophilizing the protein/antibody cation/anion pair to remove most of the water, forming a lyophilized solid; and (f) heating the lyophilized solid until a protein/antibody ionic liquid is generated. The antibody may be any desired antibody, and the anion may be any biologically-compatible anion.
1. A method for creating water-free ultra-stable antibody ionic liquid, comprising the steps of:
a) providing aqueous anti-hemoglobin antibodies;
b) cationizing aqueous antibodies by adding stoichiometric amounts of N,N-dimethyl-1,3-propanediamine (DMPDA) in the presence of a coupling reagent selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), succinimidyl iodoacetate (SIA), 2-(dimethylamino) ethanethiol, and N-(p-maleimidophenyl) isocyanate (PMPI);
c) purifying the cationized antibodies by repeated dialysis in water;
d) titrating the cationized antibodies with a counter anionic polymer of poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether (C 9 H 19 C 6 H 4 —(OCH 2 CH 2 ) 20 O(CH 2 ) 3 SO 3 ) to create at least one antibody cation/anion pair in aqueous solution until the antibody cation/anion pair solution becomes negative by zeta potential measurement;
e) dialyzing the antibodies cation/anion pair to remove excess anionic polymer using at least one molecular weight cutoff 7000 dialysis membrane;
f) lyophilizing the at least one antibody cation/anion pair to remove most of the water, forming a lyophilized solid; and
g) heating the lyophilized solid at about 27-50° C. until an antibody ionic liquid is generated.
2. The method for creating a water-free ultra-stable antibody ionic liquid of claim 1 , further comprising:
confirming the cationizing of the aqueous antibodies by measuring a positive zeta potential value.
3. The method for cretin a water-free ultra-stable antibody ionic liquid of claim 1 , further comprising:
confirming the cationizing of the aqueous antibodies by measuring a zeta potential value between about 0 and +5 mV.
4. The method for creating a water-free ultra-stable protein ionic liquid of claim 1 , further comprising:
heating the protein ionic liquid at about 100° C. for about 2 hours;
testing the antibody ionic liquid for antibody recognition of a corresponding antigen.
5. The method for creating a water-fee ultra-stable protein ionic liquid of claim 1 , further comprising:
testing the antibody ionic liquid for antibody recognition of a corresponding antigen using a dot blot assay on a nitrocellulose membrane.
6. A method for creating a water-free ultra-stable antibody ionic liquid, comprising the steps of:
a) providing aqueous anti-hemoglobin antibodies;
b) cationizing aqueous antibodies by adding stoichiometric amounts of N,N-dimethyl-1,3-propanediamine (DMPDA) in the presence of a coupling reagent selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC, succinimidyl iodoacetate (SIA), 2-(dimethylamino) ethanethiol, and N-(p-maleimidophenyl) isocyanate (PMPI),
c) purifying the cationized antibodies by repeated dialysis in water,
d) titrating the cationized antibodies with a counter anionic polymer of poly(ethylene glycol) 4-nonylphenyl 3-sulfopropyl ether (C 9 H 19 C 6 H 4 —(OCH 2 CH 2 )) 20 O(CH 2 ) 3 SO 3 ) to create at least one antibody cation/anion pair in aqueous solution until the antibody cation/anion pair solution becomes negative by zeta potential measurement;
e) dialyzing the antibodies cation/anion pair to remove excess anionic polymer using a dialysis membrane with a molecular weight cutoff of between about 6000-15,000 g/mol;
f) lyophilizing the at least one antibody cation/anion pair to remove most of the water, forming a lyophilized solid; and
g) heating the lyophilized solid at about 27-50° C. until an antibody ionic liquid is generated.