Limits...
Artificial affinity proteins as ligands of immunoglobulins.

Mouratou B, Béhar G, Pecorari F - Biomolecules (2015)

Bottom Line: A number of natural proteins are known to have affinity and specificity for immunoglobulins.Some of them are widely used as reagents for detection or capture applications, such as Protein G and Protein A.In this review, we focus on alternative scaffold proteins for which immunoglobulin binders have been identified and characterized.

View Article: PubMed Central - PubMed

Affiliation: INSERM UMR 892 - CRCNA, 8 quai Moncousu, BP 70721, 44007 Nantes Cedex 1, France. barbara.mouratou@univ-nantes.fr.

ABSTRACT
A number of natural proteins are known to have affinity and specificity for immunoglobulins. Some of them are widely used as reagents for detection or capture applications, such as Protein G and Protein A. However, these natural proteins have a defined spectrum of recognition that may not fit specific needs. With the development of combinatorial protein engineering and selection techniques, it has become possible to design artificial affinity proteins with the desired properties. These proteins, termed alternative scaffold proteins, are most often chosen for their stability, ease of engineering and cost-efficient recombinant production in bacteria. In this review, we focus on alternative scaffold proteins for which immunoglobulin binders have been identified and characterized.

Show MeSH
Some structures of molecular basis (shown in green) used to derive artificial binders with examples of associated library designs (shown in grey). (A) Synthetic domain Z based on the B domain of Staphylococcal Protein A (PDB code 1Q2N) [12] used to obtain Affibodies; (B) Sac7d protein from Sulfolobus acidocaldarius (PDB code 1AZP) [13] used to obtain Affitins; (C) Designed ankyrin repeat protein (PDB code 1MJ0) [14]; (D) Fibronectin type III domain (PDB code 1FNF) [15] used to obtain monobodies. Molecular graphics were generated using PyMOL software (The PyMOL Molecular Graphics System, Version 1.7.1.1, Schrödinger, LLC, New York, NY, USA).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4384111&req=5

biomolecules-05-00060-f001: Some structures of molecular basis (shown in green) used to derive artificial binders with examples of associated library designs (shown in grey). (A) Synthetic domain Z based on the B domain of Staphylococcal Protein A (PDB code 1Q2N) [12] used to obtain Affibodies; (B) Sac7d protein from Sulfolobus acidocaldarius (PDB code 1AZP) [13] used to obtain Affitins; (C) Designed ankyrin repeat protein (PDB code 1MJ0) [14]; (D) Fibronectin type III domain (PDB code 1FNF) [15] used to obtain monobodies. Molecular graphics were generated using PyMOL software (The PyMOL Molecular Graphics System, Version 1.7.1.1, Schrödinger, LLC, New York, NY, USA).

Mentions: Progress in the fields of molecular biology and protein engineering has led to the emergence of novel classes of tailor-made affinity proteins. A starting protein, termed an alternative scaffold protein, is often chosen to display at least the following characteristics: Small size (<20 kDa), only one polypeptide chain, high stability (thermal, chemical, etc.), high recombinant production yields and high solubility. By randomizing a set of chosen residues on the alternative scaffold protein’s surface, large libraries of variants with potentially different specificities can be created in vitro (Figure 1). Selection techniques, such as ribosome display [10] or phage display [11], can then be used to isolate from these libraries variants specific for a given target used as bait. With this approach, it is possible to generate artificial ligands with the desired properties.


Artificial affinity proteins as ligands of immunoglobulins.

Mouratou B, Béhar G, Pecorari F - Biomolecules (2015)

Some structures of molecular basis (shown in green) used to derive artificial binders with examples of associated library designs (shown in grey). (A) Synthetic domain Z based on the B domain of Staphylococcal Protein A (PDB code 1Q2N) [12] used to obtain Affibodies; (B) Sac7d protein from Sulfolobus acidocaldarius (PDB code 1AZP) [13] used to obtain Affitins; (C) Designed ankyrin repeat protein (PDB code 1MJ0) [14]; (D) Fibronectin type III domain (PDB code 1FNF) [15] used to obtain monobodies. Molecular graphics were generated using PyMOL software (The PyMOL Molecular Graphics System, Version 1.7.1.1, Schrödinger, LLC, New York, NY, USA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384111&req=5

biomolecules-05-00060-f001: Some structures of molecular basis (shown in green) used to derive artificial binders with examples of associated library designs (shown in grey). (A) Synthetic domain Z based on the B domain of Staphylococcal Protein A (PDB code 1Q2N) [12] used to obtain Affibodies; (B) Sac7d protein from Sulfolobus acidocaldarius (PDB code 1AZP) [13] used to obtain Affitins; (C) Designed ankyrin repeat protein (PDB code 1MJ0) [14]; (D) Fibronectin type III domain (PDB code 1FNF) [15] used to obtain monobodies. Molecular graphics were generated using PyMOL software (The PyMOL Molecular Graphics System, Version 1.7.1.1, Schrödinger, LLC, New York, NY, USA).
Mentions: Progress in the fields of molecular biology and protein engineering has led to the emergence of novel classes of tailor-made affinity proteins. A starting protein, termed an alternative scaffold protein, is often chosen to display at least the following characteristics: Small size (<20 kDa), only one polypeptide chain, high stability (thermal, chemical, etc.), high recombinant production yields and high solubility. By randomizing a set of chosen residues on the alternative scaffold protein’s surface, large libraries of variants with potentially different specificities can be created in vitro (Figure 1). Selection techniques, such as ribosome display [10] or phage display [11], can then be used to isolate from these libraries variants specific for a given target used as bait. With this approach, it is possible to generate artificial ligands with the desired properties.

Bottom Line: A number of natural proteins are known to have affinity and specificity for immunoglobulins.Some of them are widely used as reagents for detection or capture applications, such as Protein G and Protein A.In this review, we focus on alternative scaffold proteins for which immunoglobulin binders have been identified and characterized.

View Article: PubMed Central - PubMed

Affiliation: INSERM UMR 892 - CRCNA, 8 quai Moncousu, BP 70721, 44007 Nantes Cedex 1, France. barbara.mouratou@univ-nantes.fr.

ABSTRACT
A number of natural proteins are known to have affinity and specificity for immunoglobulins. Some of them are widely used as reagents for detection or capture applications, such as Protein G and Protein A. However, these natural proteins have a defined spectrum of recognition that may not fit specific needs. With the development of combinatorial protein engineering and selection techniques, it has become possible to design artificial affinity proteins with the desired properties. These proteins, termed alternative scaffold proteins, are most often chosen for their stability, ease of engineering and cost-efficient recombinant production in bacteria. In this review, we focus on alternative scaffold proteins for which immunoglobulin binders have been identified and characterized.

Show MeSH