Enzymatic removal of α-Gal antigen in porcine skin
10.7644/j.issn.1674-9960.2015.12.013
- VernacularTitle:酶解法清除猪皮α-Gal抗原的研究
- Author:
Zhimin YUN
;
Subo LI
;
Xue ZHANG
;
Yingxia TAN
;
Shouping JI
;
Hongwei GAO
;
Feng GONG
- Publication Type:Journal Article
- Keywords:
pig skin;
immunogenicity;
α-galactosidase;
α-Gal antigen;
residual enzyme
- From:
Military Medical Sciences
2015;39(12):938-940
- CountryChina
- Language:Chinese
-
Abstract:
Objective To reduce immunogenicity of porcine skin by removingα-Gal epitopes expressed in cell surface and extracellular matrix using recombinant α-galactosidase produced by Bacteroides fragilis.Methods The porcine skin was harvested from healthy 2-month-old pigs without any skin disorders before being sterilized by iodine and 75%alcohol, respectively.Enzymatic removal of α-Gal antigen was followed by washing with PBS.The α-Gal antigen in the prepared porcine skin was measured with immunofluorostaining of cryosections and the residual enzyme was measured with a double-antibody sandwich ELISA method.Enzymatic removal procedures were optimized by detecting residual enzyme and the effi-cacy ofα-Gal removal under different enzymatic and washing conditions.Results Efficient enzymatic and washing methods were established to removeα-Gal antigen.Theα-Gal removal efficacy was above 90% and residual enzyme was undetect-able (αprescribed minimum ofα-galactosidase detection with indirect ELISA was 1 ng/ml) .Conclusion It is feasible to efficiently removeα-Gal antigen under these enzymatic and washing conditions, and a method of producing low-immunoge-nicity pig skin dressing for burn is established.
