Assessment Method of Remnant α-1, 3-galactosyle Epitopes in Animal Tissue-derived Biomaterials.
- Author:
Yongqiang SHAN
;
Liming XU
;
Linnan KE
;
Yan LU
;
Anliang SHAO
;
Na ZHANG
;
Bixin ZENG
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
Biocompatible Materials;
Enzyme-Linked Immunosorbent Assay;
methods;
Epitopes;
analysis;
Humans;
Rats;
Sensitivity and Specificity;
Serum Albumin, Bovine;
Trisaccharides;
analysis
- From:
Journal of Biomedical Engineering
2015;32(3):662-679
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study was to establish an assessment method for determining α-Gal (α-1, 3-galactosyle) epitopes contained in animal tissue or animal tissue-derived biological materials with ELISA inhibition assay. Firstly, a 96 well plate was coated with Gal α-1, 3-Gal/bovine serum albumin (BSA) as a solid phase antigen and meanwhile, the anti-α-Gal M86 was used to react with α-Gal antigens which contained in the test materials. Then, the residual antibodies (M86) in the supernatant of M86-Gal reaction mixture were measured using ELISA inhibition assay by the α-Gal coating plate. The inhibition curve of the ELISA inhibition assay, the R2 = 0.999, was well established. Checking using both α-Gal positive materials (rat liver tissues) and α-Gal negative materials (human placenta tissues) showed a good sensitivity and specificity. Based on the presently established method, the α-Gal expression profile of rat tissues, decellular animal tissue-derived biological materials and porcine dermal before and after decellular treatment were determined. The M86 ELISA inhibition assay method, which can quantitatively determine the α-Gal antigens contained in animal tissues or animal tissue-derived biomaterials, was refined. This M86 specific antibody based-ELISA inhibition assay established in the present study has good sensitivity and specificity, and could be a useful method for determining remnant α-1, 3Gal antigens in animal tissue-derived biomaterials.
