Assessment Method of Remnantα-1, 3-galactosyle Epitopes in Animal Tissue-derived Biomaterials_Journal of Biomedical Engineering

Authors：

SHANYongqiang ^1,2 , XULiming ^1,2 , KELinnan ² , LUYan ^1,2 , SHAOAnliang ² , ZHANGNa ² ,  ZENGBixin ¹

1. School of Medical Lab Science, Wenzhou Medical University, Wenzhou 325035, China;
2. National Institutes for Food and Drug Control, Beijing 100050, China;

Corresponding author：

ZENGBixin, Email: z_bixin@163.com

Keywords：

animal tissue-derived biomaterial; α-1, 3-galactosyle epitope; ELISA inhibition assay; M86 antibody

DOI：

10.7507/1001-5515.20150120

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The aim of this study was to establish an assessment method for determiningα-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 Galα-1, 3-Gal/bovine serum albumin (BSA) as a solid phase antigen and meanwhile, the anti-α-Gal M86 was used to react withα-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 theα-Gal coating plate. The inhibition curve of the ELISA inhibition assay, the R²=0.999, was well established. Checking using bothα-Gal positive materials (rat liver tissues) andα-Gal negative materials (human placenta tissues) showed a good sensitivity and specificity. Based on the presently established method, theα-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 theα-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 remnantα-1, 3Gal antigens in animal tissue-derived biomaterials.

Citation： SHANYongqiang, XULiming, KELinnan, LUYan, SHAOAnliang, ZHANGNa, ZENGBixin. Assessment Method of Remnantα-1, 3-galactosyle Epitopes in Animal Tissue-derived Biomaterials. Journal of Biomedical Engineering, 2015, 32(3): 662-668, 679. doi: 10.7507/1001-5515.20150120 Copy

1.	BADYLAK S F, GILBERT T W. Immune response to biologic scaffold materials[J]. Semin Immunol, 2008, 20(2):109-116.
2.	BADYLAK S F. Xenogeneic extracellular matrix as a scaffold for tissue Reconstruction[J]. Transpl Immunol, 2004, 12(3/4):367-377.
3.	ALLMAN A J, MCPHERSON T B, BADYLAK S F, et al. Xenogeneic extracellular matrix grafts elicit a TH2-restricted immune response[J]. Transplantation, 2001, 71(11):1631-1640.
4.	SANDRIN M S, MCKENZIE I F. Gal alpha (1, 3)Gal, the major xenoantigen(s) recognised in pigs by human natural antibodies[J]. Immunol Rev, 1994, 141(10):169-190.
5.	JOZIASSE D H, ORIOL R. Xenotransplantation:the importance of the Galα1, 3Gal epitope in hyperacute vascular rejection[J]. Biochim Biophys Acta, 1999, 1455(2-3):403-418.
6.	COLLINS B H, COTTERELL A H, MCCURRY K R, et al. Cardiac xenografts between primate species provide evidence for the importance of the alpha-galactosyl determinant in hyperacute rejection[J]. J Immunol, 1995, 154(10):5500-5510.
7.	GALILI U, SHOHET S B, KOBRIN E, et al. Man, apes, and Old World monkeys differ from other mammals in the expression of alpha-galactosyl epitopes on nucleated cells[J]. J Biol Chem, 1988, 263(33):17755-17762.
8.	JOZIASSE D H, SHAPER N L, KIM D, et al. Murine alpha 1, 3-galactosyltransferase. A single gene locus specifies four isoforms of the enzyme by alternative splicing[J]. J Biol Chem, 1992, 267(8):5534-5541.
9.	NASO F, GANDAGLIA A, IOP L, et al. Alpha-Gal detectors in xenotransplantation research:a word of caution[J]. Xenotransplantation, 2012, 19(4):215-220.
10.	GALILI U. The alpha-gal epitope and the anti-Gal antibody in xenotransplantation and in cancer immunotherapy[J]. Immunol Cell Biol, 2005, 83(6):674-686.
11.	RIEBEN R, BOVIN N V, KORCHAGINA E Y, et al. Xenotransplantation:in vitro analysis of synthetic alpha-galactosyl inhibitors of human anti-Gal alpha 1→3Gal IgM and IgG antibodies[J]. Glycobiology, 2000, 10(2):141-148.
12.	ECKHARDTA E, GOLDSTEIN I J. Isolation and characterization of a family ofα-galactosyl containing glycoproteins from Ebrlich-ascites tumor cells[J]. Biochemistry, 1983, 22(23):5290-5297.
13.	MCKENZIE I F C, XING P X, VAUGHAN H A, et al. Distribution of the major xenoantigen (gal(α1-3)gal) for pig to human xenografts[J]. Transplant Immunology, 1994, 2(2):81-86.
14.	曾令宇, 李胜富, 张杰, 等.α-Gal表达的无创快速半定量检测[J].四川大学学报:医学版, 2005, 36(3):419-421.
15.	GALILI U, LATEMPLE D C, RADIC M Z. A sensitive assay for measuring alpha-gal epitope expression on cells by a monoclonal anti-Gal antibody[J]. Transplantation, 1998, 65(8):1129-1132.
16.	NASO F, GANDAGLIA A, LOP L, et al. First quantitative assay of alpha-Gal in soft tissues:presence and distribution of the epitope before and after cell removal from xenogeneic heart valves[J]. Acta Biomater, 2011, 7(4):1728-1734.
17.	NASO F, GANDAGLIA A, BOTTIO T, et al. First quantification of alpha-Gal epitope in current glutaraldehyde-fixed heart valve bioprostheses[J]. Xenotransplantation, 2013, 20(4):252-261.
18.	TANEMURA M, MARUYAMA S, GALILI U. Differential expression ofα-Gal epitopes (Galalpha1-3Galbeta1-4GlcNAc-R) on pig and mouse organs[J]. Transplantation, 2000, 69(1):187-190.
19.	BÖER U, LOHRENZ A, KLINGENBERG M, et al. The effect of detergent-based decellularization procedures on cellular proteins and immunogenicity in equine carotid artery grafts[J]. Biomaterials, 2011, 32(36):9730-9737.

1. BADYLAK S F, GILBERT T W. Immune response to biologic scaffold materials[J]. Semin Immunol, 2008, 20(2):109-116.
2. BADYLAK S F. Xenogeneic extracellular matrix as a scaffold for tissue Reconstruction[J]. Transpl Immunol, 2004, 12(3/4):367-377.
3. ALLMAN A J, MCPHERSON T B, BADYLAK S F, et al. Xenogeneic extracellular matrix grafts elicit a TH2-restricted immune response[J]. Transplantation, 2001, 71(11):1631-1640.
4. SANDRIN M S, MCKENZIE I F. Gal alpha (1, 3)Gal, the major xenoantigen(s) recognised in pigs by human natural antibodies[J]. Immunol Rev, 1994, 141(10):169-190.
5. JOZIASSE D H, ORIOL R. Xenotransplantation:the importance of the Galα1, 3Gal epitope in hyperacute vascular rejection[J]. Biochim Biophys Acta, 1999, 1455(2-3):403-418.
6. COLLINS B H, COTTERELL A H, MCCURRY K R, et al. Cardiac xenografts between primate species provide evidence for the importance of the alpha-galactosyl determinant in hyperacute rejection[J]. J Immunol, 1995, 154(10):5500-5510.
7. GALILI U, SHOHET S B, KOBRIN E, et al. Man, apes, and Old World monkeys differ from other mammals in the expression of alpha-galactosyl epitopes on nucleated cells[J]. J Biol Chem, 1988, 263(33):17755-17762.
8. JOZIASSE D H, SHAPER N L, KIM D, et al. Murine alpha 1, 3-galactosyltransferase. A single gene locus specifies four isoforms of the enzyme by alternative splicing[J]. J Biol Chem, 1992, 267(8):5534-5541.
9. NASO F, GANDAGLIA A, IOP L, et al. Alpha-Gal detectors in xenotransplantation research:a word of caution[J]. Xenotransplantation, 2012, 19(4):215-220.
10. GALILI U. The alpha-gal epitope and the anti-Gal antibody in xenotransplantation and in cancer immunotherapy[J]. Immunol Cell Biol, 2005, 83(6):674-686.
11. RIEBEN R, BOVIN N V, KORCHAGINA E Y, et al. Xenotransplantation:in vitro analysis of synthetic alpha-galactosyl inhibitors of human anti-Gal alpha 1→3Gal IgM and IgG antibodies[J]. Glycobiology, 2000, 10(2):141-148.
12. ECKHARDTA E, GOLDSTEIN I J. Isolation and characterization of a family ofα-galactosyl containing glycoproteins from Ebrlich-ascites tumor cells[J]. Biochemistry, 1983, 22(23):5290-5297.
13. MCKENZIE I F C, XING P X, VAUGHAN H A, et al. Distribution of the major xenoantigen (gal(α1-3)gal) for pig to human xenografts[J]. Transplant Immunology, 1994, 2(2):81-86.
14. 曾令宇, 李胜富, 张杰, 等.α-Gal表达的无创快速半定量检测[J].四川大学学报:医学版, 2005, 36(3):419-421.
15. GALILI U, LATEMPLE D C, RADIC M Z. A sensitive assay for measuring alpha-gal epitope expression on cells by a monoclonal anti-Gal antibody[J]. Transplantation, 1998, 65(8):1129-1132.
16. NASO F, GANDAGLIA A, LOP L, et al. First quantitative assay of alpha-Gal in soft tissues:presence and distribution of the epitope before and after cell removal from xenogeneic heart valves[J]. Acta Biomater, 2011, 7(4):1728-1734.
17. NASO F, GANDAGLIA A, BOTTIO T, et al. First quantification of alpha-Gal epitope in current glutaraldehyde-fixed heart valve bioprostheses[J]. Xenotransplantation, 2013, 20(4):252-261.
18. TANEMURA M, MARUYAMA S, GALILI U. Differential expression ofα-Gal epitopes (Galalpha1-3Galbeta1-4GlcNAc-R) on pig and mouse organs[J]. Transplantation, 2000, 69(1):187-190.
19. BÖER U, LOHRENZ A, KLINGENBERG M, et al. The effect of detergent-based decellularization procedures on cellular proteins and immunogenicity in equine carotid artery grafts[J]. Biomaterials, 2011, 32(36):9730-9737.

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