An efficient method for isolating antibody fragments against small peptides by antibody phage display

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt


We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of s1-casein. Key difficulties for selection of
scFv-phages against small peptides were addressed. Small peptides do not always bind efficiently to passive adsorption surfaces, and we developed a simple method to quantify the binding capacity of surfaces with the peptides. Background binding (the binding of scFvs to the background matrix) is an obstacle for successful selection, and we evaluated two methods that drastically reduced the background binding. An optimized method therefore enabled a panning procedure where the specific (peptide binding) scFv-phages were always dominant. Using 15-mer peptides immobilized on Nunc Immobilizer Streptavidin plates, we successfully generated scFvs specifically against them. The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides.

TidsskriftCombinatorial Chemistry & High Throughput Screening
Sider (fra-til)818-828
Antal sider11
StatusUdgivet - 2010


  • Det tidligere LIFE - Phage display, scFv, Tomlinson I + J libraries, casein, peptide

Antal downloads er baseret på statistik fra Google Scholar og

Ingen data tilgængelig

ID: 32447964