Establishment of a mammalian expression system for recombinant [-2]proPSA and a specific antibody against the truncated leader peptide

作者：Hwang Dobeen; Yoon Aerin; Kim Soohyun; Kim Hyori^*; Chung Junho^*

来源：Biotechnology and Applied Biochemistry, 2017, 64(3): 327-336.

DOI：10.1002/bab.1481

• [-2]proPSA
• antibody
• prostate cancer
• recombinant expression system

A truncated precursor form of prostate-specific antigen (PSA), [-2] proPSA, is a well-known biomarker for prostate cancer. To develop a biomarker assay, highly purified [-2] proPSA is required as a standard reference and for generation of a specific antibody. In this study, we generated an efficient mammalian expression system for producing a recombinant [-2] proPSA-human kappa constant domain (C.) fusion protein. N-terminal amino acid sequencing using Edman degradation demonstrated that over 95% of the recombinant protein produced is [-2] proPSA, thereby showing for the first time that recombinant [-2] proPSA can be produced as a major fraction. We also generated a recombinant chicken antibody specific to [-2] proPSA but not cross-reactive to recombinant [-7] proPSA-C., [-5] proPSA-C., and PSA purified from human seminal fluid in enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. Also, the recombinant chicken antibody reacted to recombinant [-2] proPSA protein bound to an anti-PSA antibody coated on the micrometer plate in a sandwich ELISA. All of these results suggest that the N-terminus of the [-2] proPSA-C. fusion protein resides on the exterior of the protein, thus allowing exposure to the antibody.

• 出版日期2017-6