Compositions containing HC-HA complex and methods of use thereof
Disclosed herein, in certain embodiments, is an HC•HA complex comprising hyaluronan and a heavy chain of IαI, wherein the transfer of the heavy chain of IαI is catalyzed by TSG-6. Further disclosed herein, in certain embodiments, is an HC•HA complex comprising hyaluronan and a heavy chain of IαI, wherein the transfer of the heavy chain of IαI is catalyzed by the TSG-6 like protein. Additionally, disclosed herein are methods of manufacturing said complex and methods of use thereof.
1. A method of producing a reconstituted HC•HA (rcHC•HA) complex, comprising
(a) providing HA Binding Protein (HABP) crosslinked to a stationary support;
(b) contacting the HABP crosslinked to the stationary support with
(i) hyaluronan (HA);
(ii) HC1 and HC2 of IαI; and
(iii) TSG-6; and
(c) incubating for a period of time sufficient to produce rcHC•HA complex.
2. The method of claim 1 , wherein HC1 and HC2 of IαI are isolated from serum.
3. The method of claim 1 , wherein at least one of HA, HC1, HC2, TSG-6, TSG-6-like protein is generated by a plurality of cells in the reaction mixture.
4. The method of claim 1 , further comprising isolating and purifying the rcHC•HA complex.
5. The method of claim 4 , wherein the rcHC•HA complex is isolated by centrifugation, filtration, or a combination thereof.
6. The method of claim 4 , wherein the rcHC•HA complex is purified by chromatography, gel filtration, centrifugation, differential solubility, ethanol precipitation, or a combination thereof.
7. The method of claim 6 , wherein the chromatography is affinity chromatography.