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Related Experiment Video

Updated: May 11, 2026

Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes
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Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes

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Structure-based histidine substitution for optimizing pH-sensitive Staphylococcus protein A.

Hideki Watanabe1, Hiroyuki Matsumaru, Ayako Ooishi

  • 1The National Institute of Advanced Industrial Science and Technology, Central 6, Higashi, Tsukuba 305-8566, Japan.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|May 22, 2013
PubMed
Summary
This summary is machine-generated.

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Researchers engineered a pH-sensitive Protein A variant for antibody purification. This innovation enables milder acidic conditions, reducing costs and improving efficiency in antibody manufacturing.

Area of Science:

  • Biotechnology
  • Protein Engineering
  • Biochemistry

Background:

  • Antibody purification is a costly bottleneck in antibody therapy manufacturing.
  • Current purification methods often require harsh conditions, impacting efficiency and cost.

Purpose of the Study:

  • To design a pH-sensitive Staphylococcus aureus Protein A variant for optimized antibody purification.
  • To maintain Protein A's stability and antibody affinity while introducing pH sensitivity.

Main Methods:

  • Structure-based design and mutation analysis were used to identify key substitution sites.
  • Histidine substitutions were introduced to induce electrostatic repulsion under acidic conditions.
  • The engineered variant's properties (affinity, stability, pH sensitivity) were evaluated.

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Last Updated: May 11, 2026

Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes
13:53

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Published on: December 19, 2020

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
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Published on: January 26, 2017

Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092
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Main Results:

  • Histidine substitutions decreased the dissociation rate under acidic conditions by three orders of magnitude.
  • The engineered Protein A variant demonstrated high pH sensitivity, shifting the elution peak from pH 4.2 to 5.6.
  • The variant maintained high affinity, thermal stability, and alkaline tolerance with only two substitutions.

Conclusions:

  • A novel, pH-sensitive Protein A variant was successfully engineered using a structure-based approach.
  • This variant facilitates antibody purification under milder acidic conditions, addressing manufacturing bottlenecks.
  • The strategy is applicable to engineering other protein-based ligands for various applications.