Applied Microbiology and Biotechnology

, Volume 57, Issue 4, pp 500–505

Development of novel whole-cell immunoadsorbents by yeast surface display of the IgG-binding domain

  •  Y. Nakamura
  •  S. Shibasaki
  •  M. Ueda
  •  A. Tanaka
  •  H. Fukuda
  •  A. Kondo
Original Paper

DOI: 10.1007/s002530100802

Cite this article as:
Nakamura, Y., Shibasaki, S., Ueda, M. et al. Appl Microbiol Biotechnol (2001) 57: 500. doi:10.1007/s002530100802

Abstract.

The ZZ domain derived from Staphylococcus aureus, which binds to the Fc part of immunoglobulin G (IgG), was displayed on the cell surfaces of yeast Saccharomyces cerevisiae by cell-surface engineering using the C-terminal half of α-agglutinin under control of the 5′-upstream region of the isocitrate lyase gene from Candida tropicalis (UPR-ICL). Display of ZZ on the cell surface was confirmed by immunofluorescence microscopy. Enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA using the S. cerevisiae cells displaying ZZ detected IgG and antigen (human serum albumin) down to a concentration of 1–10 ng/ml in both cases. The detection range covered by these assay systems was wide and could be varied by adjusting the amount of cells and reaction times with horseradish peroxidase (HRP) substrate. Moreover, yeast cells displaying ZZ were successfully used for repeated affinity purification of IgG from serum. These results indicate that S. cerevisiae displaying ZZ may constitute novel and genetically renewable whole-cell immunoadsorbents widely applicable to immunoassays and affinity purification.

Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  Y. Nakamura
    • 1
  •  S. Shibasaki
    • 2
  •  M. Ueda
    • 2
  •  A. Tanaka
    • 2
  •  H. Fukuda
    • 3
  •  A. Kondo
    • 1
  1. 1.Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, 1–1 Rokkodaicho, Nadaku, Kobe 657–8501, Japan
  2. 2.Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyoku, Kyoto 606–01, Japan
  3. 3.Division of Molecular Science, Graduate School of Science and Technology, Kobe-University, 1–1 Rokkodaicho, Nadaku, Kobe 657–5801, Japan