Characterization of Full-Length Recombinant Human Proteoglycan 4 as an Ocular Surface Boundary Lubricant.

Experimental eye research

PubMedID: 24997456

Samsom ML, Morrison S, Masala N, Sullivan BD, Sullivan DA, Sheardown H, Schmidt TA. Characterization of Full-Length Recombinant Human Proteoglycan 4 as an Ocular Surface Boundary Lubricant. Exp Eye Res. 2014;.
Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). Human corneas were articulated against PDMS or human eyelids, at effectively sliding velocities of 0.3-30 mm/s under physiological loads of ~15 kPa, to assess and compared the ocular lubricating ability of rhPRG4 to PRG4. Samples were tested serially in PRG4, rhPRG4 (both 300 ug/ml), then saline. Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent MW, and possessed O-linked glycosylation consistent with that of PRG4. rhPRG4 protein identity was confirmed by MS/MS. Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear.