Immunoscreening of urinary bladder cancer cDNA library and identification of potential tumor antigen

Abstract

Objective: To identify potential tumor antigen by immunoscreening the urinary bladder cancer cDNA library with monoclonal antibodies.

Methods: Monoclonal antibodies were prepared. A cDNA expression library was constructed from bladder cancer cell line BLZ211. Immunogenic proteins were identified by immunoscreening the cDNA library with ten monoclonal antibodies.

Results: The cDNA library of BLZ211 cells was established using lambdaZAP as vector. The titer of unamplified cDNA library was 1.39x10(6) pfu/ml with a recombinant rate of 97.72%, and titer of amplified one was 8.4x10(9) pfu/ml. After immunoscreening, ten positive clones representing ten different antigens were identified, which include two proteins with unknown function; coactosin-like 1, eukaryotic translation elongation factor, HNRPA1, histidine triad nucleotide binding protein, KRT7, LCN2, TSTA3, zinc finger protein, C11orf48 and HSPC148.

Conclusion: The cDNA library was of high quality and can be used in further study. By immunoscreening the bladder cancer cDNA library with ten monoclonal antibodies, we identified ten immunogenic proteins that otherwise would not have been identified as potential diagnostic marker and vaccinogens of bladder cancer using the gene discovery effort.