Method of isolation of hyaluronan
OwnerMary K. Cowman, Fok Vun Chaninventors
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Abstract
The present disclosure provides a facile method for selective isolation of hyaluronan (HA) from fluids and tissues by adsorption to a solid matrix. The method involves the use of specific solvent conditions and solid phase media favoring binding and subsequent elution of HA ranging in size from about 10 kDa to several MDa, while allowing separation of HA from other biological polyanions such as nucleic acids, sulfated glycosaminoglycans, chondroitin, and heparosan.
Claims20
A method of separating hyaluronan (HA) from a biological fluid or biological solid sample comprising:
- contacting the sample with silica in a binding solution, wherein the binding solution is an aqueous solution comprising a salt at a concentration greater than or equal to 2 M, wherein the HA binds to the silica;
- washing the silica with a washing solution to remove unbound materials;
- and
- eluting the bound HA from the silica with an elution solution,
- wherein the concentration of sulfated glycosaminoglycans of the eluted HA sample is at least 90% less than the concentration of the sulfated glycosaminoglycans of the sample prior to the contacting the sample with silica.
A method of separating hyaluronan (HA) from a biological fluid or biological solid sample comprising:
- contacting the sample with silica in a binding solution, wherein the binding solution is an aqueous solution comprising a salt at a concentration greater than or equal to 2 M, wherein the HA binds to the silica;
- washing the silica with a washing solution to remove unbound materials;
- contacting the sample with the silica in a second binding solution, wherein the second binding solution is an aqueous solution comprising a salt at a concentration greater than or equal to 4 M, wherein the HA remains bound to the silica;
- and,
- eluting the bound HA from the silica with an elution solution,
- wherein the concentration of sulfated glycosaminoglycans of the eluted HA sample is at least 90% less than the concentration of the sulfated glycosaminoglycans of the sample prior to the contacting the sample with silica.
Dependencies are inferred from the claim text.