Enhanced inhibition of human anti-gal antibody binding to mammalian cells by synthetic α-Gal epitope polymers

Jianq Qiang Wang, Xi Chen, Wei Zhang, Sima Zacharek, Yongsheng Chen, Peng George Wang

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Neoglycopolymers of polyacrylamide backbone conjugated with varying densities of Gal 1/4 -3Galβ1-4Glcβ trisaccharide epitopes (α-Gal epitopes) were designed and synthesized to study the inhibition of the binding of human natural anti-Gal antibodies to either α-Gal-containing glycoproteins or α- Gal antigens on the surface of mammalian cells. An inhibition ELISA using mouse laminin and a flow cytometry assay using pig kidney cells (PK15) were established to determine the binding affinity of the synthesized polymers. In comparison to the α-Gal monomer (Galαl-3Galβ1-4GlcNHAcβ), the α-Gal polymers dramatically enhanced the inhibition of human anti-Gal antibodies (IgG, IgM, and IgA) binding to mouse laminin or mammalian cells. Increases of 7.8 x 103- and 5.0 x 104 -fold in inhibitory potential of polymer 7C to IgA and IgM (with IC50s of 7.0 and 5.6 nM respectively) were observed over the monomer in inhibition ELISA. The results also indicated that binding enhancement of α-Gal polymers is greater for anti-Gal IgA and IgM than for IgG. Such amplified binding differences among the three anti-Gal isotypes can be utilized to selectively inhibit or remove a particular isotype of anti-Gal antibodies. Moreover, it was demonstrated through the flow cytometry assay that certain α-Gal polymers are effective in inhibition of anti-Gal antibody (in human serum) binding to pig kidney (PK15) cells. Thus, such synthetic carbohydrate polymers may find practical applications in cell xenotransplantations.

Original languageEnglish (US)
Pages (from-to)8174-8181
Number of pages8
JournalJournal of the American Chemical Society
Volume121
Issue number36
DOIs
StatePublished - Sep 15 1999
Externally publishedYes

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Artificial Cells
Epitopes
Antibodies
Anti-Idiotypic Antibodies
Polymers
Cells
Immunoglobulin A
Immunoglobulin M
Flow cytometry
Laminin
Assays
Flow Cytometry
Swine
Immunoglobulin G
Monomers
Enzyme-Linked Immunosorbent Assay
Kidney
Trisaccharides
Heterologous Transplantation
Glycoproteins

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Enhanced inhibition of human anti-gal antibody binding to mammalian cells by synthetic α-Gal epitope polymers. / Wang, Jianq Qiang; Chen, Xi; Zhang, Wei; Zacharek, Sima; Chen, Yongsheng; Wang, Peng George.

In: Journal of the American Chemical Society, Vol. 121, No. 36, 15.09.1999, p. 8174-8181.

Research output: Contribution to journalArticle

Wang, Jianq Qiang ; Chen, Xi ; Zhang, Wei ; Zacharek, Sima ; Chen, Yongsheng ; Wang, Peng George. / Enhanced inhibition of human anti-gal antibody binding to mammalian cells by synthetic α-Gal epitope polymers. In: Journal of the American Chemical Society. 1999 ; Vol. 121, No. 36. pp. 8174-8181.
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abstract = "Neoglycopolymers of polyacrylamide backbone conjugated with varying densities of Gal 1/4 -3Galβ1-4Glcβ trisaccharide epitopes (α-Gal epitopes) were designed and synthesized to study the inhibition of the binding of human natural anti-Gal antibodies to either α-Gal-containing glycoproteins or α- Gal antigens on the surface of mammalian cells. An inhibition ELISA using mouse laminin and a flow cytometry assay using pig kidney cells (PK15) were established to determine the binding affinity of the synthesized polymers. In comparison to the α-Gal monomer (Galαl-3Galβ1-4GlcNHAcβ), the α-Gal polymers dramatically enhanced the inhibition of human anti-Gal antibodies (IgG, IgM, and IgA) binding to mouse laminin or mammalian cells. Increases of 7.8 x 103- and 5.0 x 104 -fold in inhibitory potential of polymer 7C to IgA and IgM (with IC50s of 7.0 and 5.6 nM respectively) were observed over the monomer in inhibition ELISA. The results also indicated that binding enhancement of α-Gal polymers is greater for anti-Gal IgA and IgM than for IgG. Such amplified binding differences among the three anti-Gal isotypes can be utilized to selectively inhibit or remove a particular isotype of anti-Gal antibodies. Moreover, it was demonstrated through the flow cytometry assay that certain α-Gal polymers are effective in inhibition of anti-Gal antibody (in human serum) binding to pig kidney (PK15) cells. Thus, such synthetic carbohydrate polymers may find practical applications in cell xenotransplantations.",
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