Identification of tumor-associated antigens by screening phage-displayed human cDNA libraries with sera from tumor patients.
Screening cDNA libraries from solid human tumors with sera of autologous patients (SEREX) has proven to be a powerful approach to identifying tumor antigens recognized by the humoral arm of the immune system. In many cases, application of this methodology has led to the discovery of novel tumor antigens as unknown gene products. We tried to improve the potency of the SEREX approach by combining it with phage-display technology. We designed a new lambda vector to express protein fragments as N-terminal fusions to the D capsid protein and generated high-complexity cDNA libraries from human breast carcinoma cell lines and solid tumors. Screening these phage-displayed libraries required limited amounts of sera from patients and efficiently identified several tumor antigens specifically reacting with sera from breast cancer patients.
Minenkova
Olga
O
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Pucci
Andrea
A
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Pavoni
Emiliano
E
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
De Tomassi
Amedeo
A
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Fortugno
Paola
P
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Gargano
Nicola
N
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Cianfriglia
Maurizio
M
Laboratorio di Immunologia, Reparto Immunologia dei Tumori, Istituto Superiore di Sanità, Roma, Italy.
Barca
Stefano
S
Laboratorio di Immunologia, Reparto Immunologia dei Tumori, Istituto Superiore di Sanità, Roma, Italy.
De Placido
Sabino
S
Dipartimento di Endocrinologia ed Oncologia Molecolare e Clinica, Università di Napoli Federico II, Napoli, Italy.
Martignetti
Angelo
A
Dipartimento di Endocrinologia ed Oncologia Molecolare e Clinica, Università di Napoli Federico II, Napoli, Italy.
Felici
Franco
F
Kenton Labs, c/o Sigma Tau, Pomezia (Roma), Italy.
Cortese
Riccardo
R
Istituto di Ricerche di Biologia Molecolare P. Angeletti, Pomezia (Roma), Italy.
Monaci
Paolo
P
Istituto di Ricerche di Biologia Molecolare P. Angeletti, Pomezia (Roma), Italy.
D000328:Adult; D000368:Aged; D000818:Animals; D000912:Antibodies, Neoplasm / Q000097:blood; D000951:Antigens, Neoplasm / Q000032:analysis*; D017104:Bacteriophage M13 / Q000235:genetics; D010582:Bacteriophage lambda; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D004273:DNA, Neoplasm / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D015723:Gene Library; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008297:Male; D008403:Mass Screening; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D012333:RNA, Messenger / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured
D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D017931:DNA Primers; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D005982:Glutathione Transferase