Production and radioimmunoimaging of novel fully human phage display recombinant antibodies and growth inhibition of lung adenocarcinoma cell line overexpressing Prx I.
The Peroxiredoxin I (Prx I) is a member of the Peroxiredoxin family, which is overexpressed in many diverse tumor types and is an anti-apoptosis protein for tumor cell proliferation and survival. Therapeutic strategies targeting the Prx I may therefore be effective broad-spectrum anticancer agents. We constructed a phage display single-chain variable fragment (scFv) antibody library and sieve out the fully human, lung adenocarcinoma-sepcific monoclonal antibodies. The selection on Prx I was performed using above-mentioned lung adenocarcinoma-sepcific monoclonal antibodies with high affinity to Prx I overexpressing lung adenocarcinoma cells. The candidate scFv sequences, based on enzyme-linked immunosorbent assay (ELISA) screening data, were chosen for soluble expression, and a 30 kDa band was observed on polyacrylamide gel electrophoresis as predicted. The purified antibodies were characterized by immunoblotting and showed high specificity to Prx I-overexpressing lung adenocarcinoma cells A549. Radioimmunoimaging was taken to evaluate specificity and distribution of antibodies in vivo. The radiolocalization index (RI) of tumor/serum and tumor/muscle gradually increased, reaching its peak (4.06 +/- 0.13 and 5.17 +/- 0.97, respectively) at 48 h postadministration. Single photon emission computed tomography (SPECT) imaging showed the radioactivity was aggregated in tumor locations and tumor imaging was clearly observed. The internalized scFv resulted in antibody-mediated cell apoptosis and downregulation of Prx I expression. These results demonstrate that the scFv possesses strong antitumor activity on lung adenocarcinoma and may therefore be an effective therapeutic candidate for the treatment of cancers that are dependent on Prx I for growth and survival.
Luo
Yi
Y
Department of Radiology, Chongqing Medical University, Chongqing, China.
Pang
Hua
H
Li
Shujie
S
Cao
Hui
H
Peng
Zhiping
Z
Fan
Chunbo
C
Li
Shaolin
S
D000230:Adenocarcinoma / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D000818:Animals; D000915:Antibody Affinity; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D001980:Bronchi / Q000166:cytology / Q000276:immunology; D016503:Drug Delivery Systems; D004847:Epithelial Cells / Q000276:immunology; D005803:Genes, Immunoglobulin; D006801:Humans; D007457:Iodine Radioisotopes; D008175:Lung Neoplasms / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology; D019151:Peptide Library; D054464:Peroxiredoxins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology; D016719:Radioimmunodetection; D011994:Recombinant Proteins / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology / Q000302:isolation & purification / Q000627:therapeutic use*; D014018:Tissue Distribution; D015899:Tomography, Emission-Computed, Single-Photon; D023041:Xenograft Model Antitumor Assays
D007457:Iodine Radioisotopes; D009363:Neoplasm Proteins; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C515940:PRDX1 protein, human; D054464:Peroxiredoxins