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Related Experiment Video

Updated: May 28, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
10:32

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

Published on: January 16, 2012

Engineering bispecificity into a single albumin-binding domain.

Johan Nilvebrant1, Tove Alm, Sophia Hober

  • 1Department of Proteomics, School of Biotechnology, Royal Institute of Technology, AlbaNova University Center, Stockholm, Sweden.

Plos One
|October 13, 2011
PubMed
Summary

Researchers engineered ultra-small bispecific proteins, just 46 amino acids, targeting tumor necrosis factor-alpha (TNF-α) and albumin. These novel binders demonstrate high affinity, paving the way for advanced biotherapeutics.

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Area of Science:

  • Biotechnology
  • Protein Engineering
  • Immunotherapy

Background:

  • Bispecific antibodies and proteins offer significant potential in biotherapeutics.
  • Developing smaller, highly effective bispecific proteins remains a key challenge.

Purpose of the Study:

  • To engineer novel, extremely small bispecific proteins using a single structural domain.
  • To achieve high-affinity binding to both tumor necrosis factor-alpha (TNF-α) and albumin.

Main Methods:

  • Engineered an albumin-binding domain to also bind TNF-α, creating a 46-amino acid bispecific protein.
  • Utilized phage display and staphylococcal surface display for library diversification, selection, and affinity maturation.
  • Employed multiparameter sorting targeting both TNF-α and albumin simultaneously.

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A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
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A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity

Published on: July 12, 2018

Related Experiment Videos

Last Updated: May 28, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
10:32

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

Published on: January 16, 2012

A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
11:13

A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity

Published on: July 12, 2018

Main Results:

  • Successfully generated bispecific single-domain proteins with high affinity for TNF-α (below 5 nM).
  • Isolated variants with high affinity for both TNF-α and albumin using simultaneous multiparameter sorting.
  • Developed the smallest engineered bispecific proteins reported to date.

Conclusions:

  • The novel strategy enables the efficient engineering of ultra-small, high-affinity bispecific proteins.
  • Cell display technologies are suitable for developing bispecificity with multiple targets.
  • These findings open new avenues for biotherapeutic applications with enhanced specificity and reduced size.