Profiling the autoantibody repertoire by screening phage-displayed human cDNA libraries.
The advent of the serological identification of antigens by procedures such as cDNA cloning and recombinant protein expression has allowed the direct molecular definition of immunogenic proteins. The phage-display technology provides several advantages over conventional immunoscreening procedures based on plasmid or lambda-phage cDNA libraries. So far, attempts to display open reading frames, such as those encoded by cDNA fragments, on filamentous phages have not been very successful. We managed to develop a strategy based on "folding reporters" which allows filtering out open reading frames from DNA and displaying them on filamentous phages in such a way that they are amenable to subsequent selection or screening. Once the cDNA library of interest is created, phage-display technology is used for selection of novel putative antigens; these are then validated by printing isolated protein on microarray and screening with patients' sera.
Di Niro
Roberto
R
Department of Life Sciences, University of Trieste, Trieste, Italy.
D'Angelo
Sara
S
Secco
Paola
P
Marzari
Roberto
R
Santoro
Claudio
C
Sblattero
Daniele
D
D001323:Autoantibodies / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008954:Models, Biological; D016366:Open Reading Frames; D019151:Peptide Library; D040081:Protein Array Analysis / Q000379:methods; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology
D001323:Autoantibodies; D019151:Peptide Library; D011994:Recombinant Proteins