BIOMAT Database Logo BIOMAT Database Logo

Development and Characterization of Peptide Ligands of Immunoglobulin G Fc Region. 2018

Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
University of Ljubljana , Faculty of Pharmacy, Department of Pharmaceutical Biology , SI-1000 Ljubljana , Slovenia.

Affinity chromatography based on bacterial immunoglobulin (Ig)-binding proteins represents the cornerstone of therapeutic antibody downstream processing. However, there is a pressing need for more robust affinity ligands that would withstand the harsh column sanitization conditions, while still displaying high selectivity for antibodies. Here, we report the development of linear peptide IgG ligands, identified from combinatorial phage-display library screens. The lead peptide was shown to compete with staphylococcal protein A for the IgG Fc region. Trimming analysis and alanine scanning revealed the minimal structural requirements of the peptide for Fc binding, and the minimized peptide GSYWYQVWF recognized all human IgG subtypes. Mutation of glutamine located at the nonessential position 6 to aspartate led to the optimized peptide GSYWYDVWF with 18-fold higher affinity ( KD app. 0.6 μM) compared to the parent peptide. When coupled to paramagnetic beads or a chromatographic matrix, the optimized ligand was shown to selectively enrich antibodies from complex protein mixtures.

UI MeSH Term Description Entries
D007074 Immunoglobulin G The major immunoglobulin isotype class in normal human serum. There are several isotype subclasses of IgG, for example, IgG1, IgG2A, and IgG2B. Gamma Globulin, 7S,IgG,IgG Antibody,Allerglobuline,IgG(T),IgG1,IgG2,IgG2A,IgG2B,IgG3,IgG4,Immunoglobulin GT,Polyglobin,7S Gamma Globulin,Antibody, IgG,GT, Immunoglobulin
D007141 Immunoglobulin Fc Fragments Crystallizable fragments composed of the carboxy-terminal halves of both IMMUNOGLOBULIN HEAVY CHAINS linked to each other by disulfide bonds. Fc fragments contain the carboxy-terminal parts of the heavy chain constant regions that are responsible for the effector functions of an immunoglobulin (COMPLEMENT fixation, binding to the cell membrane via FC RECEPTORS, and placental transport). This fragment can be obtained by digestion of immunoglobulins with the proteolytic enzyme PAPAIN. Fc Fragment,Fc Fragments,Fc Immunoglobulin,Fc Immunoglobulins,Ig Fc Fragments,Immunoglobulin Fc Fragment,Immunoglobulins, Fc,Immunoglobulins, Fc Fragment,Fc Fragment Immunoglobulins,Fc Fragment, Immunoglobulin,Fc Fragments, Ig,Fc Fragments, Immunoglobulin,Fragment Immunoglobulins, Fc,Fragment, Fc,Fragments, Ig Fc,Immunoglobulin, Fc
D008024 Ligands A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed) Ligand
D010455 Peptides Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are considered to be larger versions of peptides that can form into complex structures such as ENZYMES and RECEPTORS. Peptide,Polypeptide,Polypeptides
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001435 Bacteriophages Viruses whose hosts are bacterial cells. Phages,Bacteriophage,Phage
D013205 Staphylococcal Protein A A protein present in the cell wall of most Staphylococcus aureus strains. The protein selectively binds to the Fc region of human normal and myeloma-derived IMMUNOGLOBULIN G. It elicits antibody activity and may cause hypersensitivity reactions due to histamine release; has also been used as cell surface antigen marker and in the clinical assessment of B lymphocyte function. Protein A,Protein A, Staphylococcal

Related Publications

Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Immunochemical study of human immunoglobulin G Fc region.
February 1996, Cancer biotherapy & radiopharmaceuticals,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Genetic and antigenic analysis of mink's immunoglobulin G Fc region.
January 2014, Central-European journal of immunology,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Pairing of oligosaccharides in the Fc region of immunoglobulin G.
May 2000, FEBS letters,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
The functional differentiation of the Fc region of immunoglobulin G.
January 1985, Annales de l'Institut Pasteur. Immunologie,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Structural characterization of anti-inflammatory immunoglobulin G Fc proteins.
September 2014, Journal of molecular biology,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Cloning and characterization of ovine immunoglobulin G Fc receptor III (FcγRIII).
February 2011, Veterinary immunology and immunopathology,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Cloning and characterization of ovine immunoglobulin G Fc receptor II (FcgammaRII).
February 2010, Veterinary immunology and immunopathology,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Fc-Binding Ligands of Immunoglobulin G: An Overview of High Affinity Proteins and Peptides.
December 2016, Materials (Basel, Switzerland),
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
Identification and characterization of human immunoglobulin G Fc receptors of Fusobacterium nucleatum.
April 2000, Oral microbiology and immunology,
Nika Kruljec, and Peter Molek, and Vesna Hodnik, and Gregor Anderluh, and Tomaž Bratkovič
RNA aptamers specifically interact with the Fc region of mouse immunoglobulin G.
January 2008, Nucleic acids symposium series (2004),
Copied contents to your clipboard!