Engineered High Affinity Antibodies for Detection of Phosphotyrosine Proteins

Tech ID: 29924 / UC Case 2018-237-0

Invention Novelty

This invention comprises high affinity phosphotyrosine antibodies with greater thermal stability and detection capability for Western Blotting and Immunofluorescence compared to popular commercial antibodies.

Value Proposition

Phosphotyrosine (pY) is one of the most important posttranslational modifications that regulate many vital human signaling pathways. Dysregulation of pY modifications leads to various diseases including neurodegenerative disorders and cancer. pY antibodies have immerged as powerful tools to understand such modifications in the past two decades. There are several commercially available pan-specific pY antibodies. 4G10 is the market leader in this category but many low-abundance pY proteins or proteins in specific molecular weight ranges are difficult to detect by this and other pY antibodies. Engineering pY antibodies to improve its affinity would enable better detection of low abundance pY molecules in the cell.

Our researchers have created several affinity-improved pY antibodies that outperform the popular commercial pY antibody 4G10 in both Western Blotting and immunofluorescence. These improved antibodies can be used for flow cytometry, immunoprecipitation, pY proteomics and other pY detection technologies.

Engineered antibodies have the following advantages as compared to 4G10:

  • Detects additional low-abundance proteins in western blots
  • Provides ~ 200-fold brighter immunofluorescence signals
  • High thermal stability with Tm > 70°C for long-term storage
  • Small-scale production shows improved protein yields

Technology Description

Researchers at University of California, San Francisco (UCSF) have engineered pY antibodies that display higher affinity, detection capability and thermal stability than any commercially available pan-specific pY antibodies. First, they stabilized the 4G10 antibody by grafting its complementarity-determining regions to a highly thermo-stable human scaffold. Then they successfully solved the crystal structure of this stabilized scaffold complex with a pY substrate. Based on this structure, they rationally designed and generated improved pY antibodies by protein engineering.

Stage of Development

Proof of Concept

Related Materials

  • The publication is in press.

Data Availability

In vitro data

Patent Status

Patent Pending

Patent Status

Provisional application is filed

Contact

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Inventors

  • Mou, Yun
  • Wells, James (Jim) A.

Other Information

Keywords

Phosphotyrosine, Antibody engineering

Categorized As