Pular para o conteúdo
Merck
  • High-throughput screening of glycan-binding proteins using miniature pig kidney N-glycan-immobilized beads.

High-throughput screening of glycan-binding proteins using miniature pig kidney N-glycan-immobilized beads.

Chemistry & biology (2008-03-22)
Yun-Gon Kim, Dong-Sik Shin, Yung-Hun Yang, Geun-Cheol Gil, Chung-Gyu Park, Yusuke Mimura, David K C Cooper, Pauline M Rudd, Raymond A Dwek, Yoon-Sik Lee, Byung-Gee Kim
RESUMO

Glycan recognition leading to cell-cell interactions, signaling, and immune responses is mediated by various glycan-binding proteins (GBPs) showing highly diverse ligand specificities. We describe here a rapid glycan immobilization technique via 4-hydrazinobenzoic acid (HBA)-functionalized beads and its application to high-throughput screening of miniature pig kidney N-glycan-binding proteins by using a mass-spectrometric approach. Without any derivatization steps, the purified pig kidney N-glycans were directly immobilized on to HBA-functionalized beads and subsequently used to identify GBPs from human serum. This screening method showed remarkable performance for identifying potential GBPs closely involved in pig-to-human xenograft rejection mediated by human serum, including antibodies, cytokines, complement components, siglec, and CD antigens. Thus, these results demonstrate that the GBP screening method was firmly established by one-step immobilization of the N-glycans on to microsphere and highly sensitive mass-spectrometric analysis.

MATERIAIS
Número do produto
Marca
Descrição do produto

Sigma-Aldrich
4-Hydrazinobenzoic acid, 97%
Sigma-Aldrich
4-Hydrazinobenzoic acid hydrochloride, 98%