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Identification of a ligand for IgG‐Fc derived from a soluble peptide library based on fusion proteins secreted by S. cerevisiae
Author(s) -
Mersich Christa,
Billes Werner,
Jungbauer Alois
Publication year - 2007
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200700049
Subject(s) - peptide , peptide library , computational biology , ligand (biochemistry) , monoclonal antibody , fusion protein , identification (biology) , high throughput screening , chemistry , biochemistry , antibody , combinatorial chemistry , biology , microbiology and biotechnology , peptide sequence , recombinant dna , receptor , immunology , gene , botany
Biological libraries are important tools in the development of new peptide‐based compounds. Here, we describe the use of a soluble peptide library system as a complementary tool in the field of ligand development. Random peptides were expressed in S. cerevisiae as carboxy‐terminal extensions of the eukaryotic initiation factor 5a (eIF5a) and secreted into the culture supernatant. Expression and screening of this library were performed in a microwell format. As an example of this versatile approach, we describe the identification of a ligand for the human IgG‐Fc fragment. Ligands binding IgG‐Fc show great potential in a wide variety of applications including development of therapeutics, streamlining the large‐scale purification of antibodies, and applications in diagnostic tests. We demonstrated the utility of this system. After screening only 6160 clones, we identified a ligand with the peptide sequence of TRRRTCSPPTWPRARARSTPSGCSSTGPSANRG. An affinity constant of 3.9 x 10 5 M ‐1 was determined by a biosensor method. Handling and maintenance of this library is conceptually simple and highly applicable for automated high‐throughput systems.