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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

12 Issues per year


IMPACT FACTOR 2016: 3.273

CiteScore 2016: 3.01

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1437-4315
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Volume 387, Issue 6 (Jun 2006)

Issues

Human tissue kallikrein 9: production of recombinant proteins and specific antibodies

Nader Memari
  • Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto M5G 1X5, ON, Canada and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto M5G 1L5, ON, Canada
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Linda Grass
  • Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto M5G 1X5, ON, Canada
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Terukazu Nakamura
  • Department of Urology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kawaramachi-Hirokoji, Kyoto 602-8566, Japan
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Inci Karakucuk
  • Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto M5G 1X5, ON, Canada
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Eleftherios P. Diamandis
  • Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto M5G 1X5, ON, Canada and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto M5G 1L5, ON, Canada
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2006-06-26 | DOI: https://doi.org/10.1515/BC.2006.092

Abstract

Human tissue kallikreins (genes, KLKs; proteins, hKs) are a subgroup of hormonally regulated serine proteases. Two tissue kallikreins, namely hK2 and hK3 (prostate-specific antigen, PSA), are currently used as serological biomarkers of prostate cancer. Human tissue kallikrein 9 (KLK9) is a newly identified member of the tissue kallikrein gene family. Recent reports have indicated that KLK9 mRNA is differentially expressed in ovarian and breast cancer and has prognostic value. Here, we report the production of recombinant hK9 (classic form) using prokaryotic and mammalian cells and the generation of polyclonal antibodies. Total testis tissue mRNA was reverse-transcribed to cDNA, amplified, cloned into a pET/200 TOPO plasmid vector, and transformed into E. coli cells. hK9 was purified and used as an immunogen to generate polyclonal antibodies. Full-length KLK9 cDNA was also cloned in the vector pcDNA3.1 and was expressed in CHO cells. The identity of hK9 was confirmed by mass spectrometry. hK9 rabbit antiserum displayed no cross-reactivity with other tissue kallikreins and could specifically recognize E. coli- and CHO-derived hK9 on Western blots. hK9 was mainly detected in testis and seminal vesicles by Western blotting. The reagents generated here will help to define the physiological role of this tissue kallikrein and its involvement in human disease.

Keywords: antibody generation; cancer biomarkers; human tissue kallikreins; protein expression; protein purification; serine proteases

About the article

Corresponding author


Received: October 28, 2005

Accepted: February 15, 2006

Published Online: 2006-06-26

Published in Print: 2006-06-01


Citation Information: Biological Chemistry, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2006.092.

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