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Cell Surface Display Systems for Protein Engineering

Creative Biolabs has adopted an important technology for protein engineering and design effectively -- the Cell Surface Display Systems.

Cell surface display systems are used to express a protein or peptide on the cellular surface of prokaryotes and eukaryotes (e.g., bacteria, yeast, insect and mammalian cells). These systems have been used for many different biotechnical and biomedical applications, proving their effectiveness in protein engineering studies, such as vaccine development, bioabsorbants, biocatalysts, biosensors and so on. Compared with other display strategies, cell surface display systems have the following advantages of being:

  1. Freely accessible and stable for the binder bound with the target;
  2. Easily analyzable by high-throughput methods like Fluorescence-Activated Cell Sorting (FACS), magnetic bead sorting and panning because of their large sizes of bacterial and yeast cells;
  3. Quantitative in flow cytometry screening at a rate of > 104 clones/sec and able to eliminate the random bias when small proteins or peptides are displayed;
  4. Simple in operation by low-speed centrifugations during the library screening step when compared to phages.

The displayed proteins or peptides with distinct functions at the cellular surface freely are accessible for the substrate or binding partner in activity or binding studies. Besides, with the cell envelope as a matrix, the proteins or peptides have proven to be more stable, thus making associated preparation or purification redundant in many instances.

It has been recently identified that glycosylation significantly enhances epitope presentation. However, prokaryotic expression systems have been conventionally used as the predominant way of producing therapeutic proteins fused with cell-wall anchoring motifs. To solve this, L. lactis was used to surface display the glycosylated murine Tyrosinase related protein-2 (TRP-2), which was fused with cell wall anchoring LysM motif (cA) at the C-terminus. The success also implies possible development of novel and inexpensive vaccine platforms.

Yeast surface display has also been used to screen a combinatorial heterodimeric Fc library. Heterodimeric Fc variants were first generated through directed evolution combined and then displayed using yeast haploid cells. Heterodimerization took place in the mated cells, and its detection would enable the screening of the Fc library. This provides a new approach for engineering not only Fc heterodimers but also other protein heterodimers via directed evolution combined with yeast surface display.


Figure 1. The screening of cell-based libraries using FCM. (Patrick S. Daugherty et al., 2000)

There are still other applications for cell surface display systems. And we can work with you on your protein engineering projects. So contact us and find out more.

Reference

  1. Patrick S. Daugherty, Brent L. Iverson. Flow cytometric screening of cell-based libraries. Journal of Immunological Methods 2000, 243: 211–227.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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