ArticleJournal of immunological methods2014
Applications for an engineered Protein-G variant with a pH controllable affinity to antibody fragments.
Article in Journal of immunological methods, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 40 citations in OpenAlex.
- Overcoming the Limitations of Protein A: Evolution of Bacterial Protein-Based and Synthetic Affinity Ligands for High-Performance IgG Purification.International journal of molecular sciences · 2026Review
- A universal Fab targeting a conserved U1A-RNA epitope for RNA structure determination by cryo-EM.Nucleic acids research · 2026Article
- Engineering of an Fc-specific monovalent protein G for the light-controlled affinity purification of antibodies.Scientific reports · 2025Article
- A universal chimeric antigen receptor (CAR)-fragment antibody binder (FAB) split system for cancer immunotherapy.Science advances · 2025Article
- Integrating Affinity Chromatography in the Platform Process for Adenovirus Purification.Biotechnology and bioengineering · 2025Article
- Engineering and Structural Elucidation of a Sac7d-Derived IgG Fc-Specific Affitin and Its Application for the Light-Controlled Affinity Purification of Antibodies.Chembiochem : a European journal of chemical biology · 2025Article
- Stomata-targeted nanocarriers enhance plant defense against pathogen colonization.Nature communications · 2025Article
- Engineered protein G variants for multifunctional antibody-based assemblies.Protein science : a publication of the Protein Society · 2025Article
- Neural network extrapolation to distant regions of the protein fitness landscape.Nature communications · 2024Article
- [Recent progress of chromatographic techniques for antibody purification].Se pu = Chinese journal of chromatography · 2024Review
- A Generic Antibody-Blocking Protein That Enables pH-Switchable Activation of Antibody Activity.ACS chemical biology · 2024Article
- Neural network extrapolation to distant regions of the protein fitness landscape.bioRxiv : the preprint server for biology · 2023Article
- Article
- Engineering of a synthetic antibody fragment for structural and functional studies of K+ channels.The Journal of general physiology · 2022Article
- Protein Engineering: Advances in Phage Display for Basic Science and Medical Research.Biochemistry. Biokhimiia · 2022Review
- Engineered Ultra-High Affinity Synthetic Antibodies for SARS-CoV-2 Neutralization and Detection.Journal of molecular biology · 2021Article
- Recognition of an α-helical hairpin in P22 large terminase by a synthetic antibody fragment.Acta crystallographica. Section D, Structural biology · 2020Article
- An engineered ultra-high affinity Fab-Protein G pair enables a modular antibody platform with multifunctional capability.Protein science : a publication of the Protein Society · 2020Article
- Development of "Plug and Play" Fiducial Marks for Structural Studies of GPCR Signaling Complexes by Single-Particle Cryo-EM.Structure (London, England : 1993) · 2019Article
- Rapid Discovery and Characterization of Synthetic Neutralizing Antibodies against Anthrax Edema Toxin.Biochemistry · 2019Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
Immunoglobulin binding proteins (IBPs) are broadly used as reagents for the purification and detection of antibodies. Among the IBPs, the most widely used are Protein-A and Protein-G. The C2 domain of Protein-G from Streptococcus is a multi-specific protein domain; it possesses a high affinity (K(D) ~10 nM) for the Fc region of the IgG, but a much lower affinity (KD~low μM) for the constant domain of the antibody fragment (Fab), which limits some of its applications. Here, we describe the engineering of the Protein-G interface using phage display to create Protein-G-A1, a variant with 8 point mutations and an approximately 100-fold improved affinity over the parent domain for the 4D5 Fab scaffold. Protein-G-A1 is capable of robust binding to Fab fragments for numerous applications. Furthermore, we isolated a variant with pH-dependent affinity, demonstrating a 1,000-fold change in affinity from pH7 to 4. Additional rational mutagenesis endowed Protein-G with significantly enhanced stability in basic conditions relative to the parent domain while maintaining high affinity to the Fab. This property is particularly useful to regenerate Protein-G affinity columns. Lastly, the affinity-matured Protein-G-A1 variant was tethered together to produce dimers capable of providing multivalent affinity enhancement to a low affinity antibody fragment-antigen interaction. Engineered Protein-G variants should find widespread application in the use of Fab-based affinity reagents.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.