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Published on: September 17, 2014
[Purification of corneal endothelial antigen and anti-corneal endothelial antibody]
1Department of Ophthalmology, First Military Medical University, Guangzhou 510515, China.
Objective:
To explore the methods for purifying corneal endothelial antigen (CEN-Ag) and preparing anti-corneal endothelial antibody (aCEN-Ab).
Methods:
Porcine corneal endothelia were obtained with the epithelia serving as control, CEN-Ag and corneal epithelial antigen (CEP-Ag) were isolated and purified by ion-exchange chromatography and affinity chromatography with 2B4.14.1 monoclonal antibody. The rabbits were immunized with the resultant CEN-Ag and CEP-Ag and their sera collected to harvest aCEN-Ab and anti-corneal epithelial antibody (aCEP-Ab) by affinity chromatography.
Results And Conclusion:
The relative molecular mass of CEN-Ag and CEP-Ag were 43 000 and 53 000 respectively as determined by SDS-PAGE, with their immunoglobulins being IgG2 and IgG3.
