Identification of prostate cancer antigens by automated high-throughput filter immunoscreening.
There is a need for earlier and more accurate cancer diagnostics as well as new targets for cancer immunotherapy. To this end, it is important to identify sets of tumour antigens specific for different cancer forms. Several methods that identify potential tumour antigens in an arrayed and high-throughput format have been developed during the last years of SEREX (serological identification of antigens by recombinant expression cloning) related research. Such techniques may hold the potential to describe the complete immunogenic part of the cancer proteome, also called the cancer immunoproteome. We have developed a powerful platform for automated serological high-throughput filter screening of tumour cDNA libraries. The screening format of this method is 18,000 single cDNAs clones, which is superior to other high-throughput methods described. The output is antigens, which are potential diagnostic cancer markers and vaccine targets. We present here the results from the screening of a prostate tumour cDNA library with autologous patient antibodies. We first demonstrated the feasibility of the automated high-throughput filter immunoscreening method by use of the NY-ESO-1sv (NY-ESO-1 splice variant) antigen. We then screened 18,000 cDNA clones from a phage display selected prostate tumour cDNA library with autologous patient antibodies and identified several relevant antigens such as NY-ESO-1, XAGE-1, DJ-1 and transcription factor 25 (TCF25). The present high-throughput immunoscreening method has the potential to identify both patient-specific and disease-specific antigens for use in diagnostics and therapy.
Department of Immunology, Institute for Cancer Research, Rikshospitalet-Radiumhospitalet Medical Centre, Montebello, Oslo, Norway. lene.alsoe@rr-research.no
Lene
L
Alsøe
John Erik
JE
Stacy
Alexander
A
Fosså
Steinar
S
Funderud
Ole Henrik
OH
Brekke
Gustav
G
Gaudernack
D000912:Antibodies, Neoplasm / Q000097:blood*; D000937:Antigen-Antibody Reactions; D000951:Antigens, Neoplasm / Q000032:analysis* / Q000235:genetics; D001331:Automation; D051792:Basic Helix-Loop-Helix Proteins / Q000032:analysis; D003001:Cloning, Molecular; D003101:Collodion; D015252:Deoxyribonucleases, Type II Site-Specific; D005240:Feasibility Studies; D015723:Gene Library; D006801:Humans; D015151:Immunoblotting / Q000295:instrumentation; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis; D008198:Lymph Nodes / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis; D008567:Membranes, Artificial; D015513:Oncogene Proteins / Q000032:analysis; D011471:Prostatic Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000276:immunology*; D000071617:Protein Deglycase DJ-1; D012097:Repressor Proteins / Q000032:analysis; D015203:Reproducibility of Results; D015183:Restriction Mapping
D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D051792:Basic Helix-Loop-Helix Proteins; D003101:Collodion; D015252:Deoxyribonucleases, Type II Site-Specific; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008567:Membranes, Artificial; D015513:Oncogene Proteins; D000071617:Protein Deglycase DJ-1; D012097:Repressor Proteins; C104948:CTAG1B protein, human; C510400:TCF25 protein, human; C413656:XAGE1A protein, human; C105131:PARK7 protein, human; C052380:endodeoxyribonuclease NcoI