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Development and evaluation of an ELISA method for the measurement of kallikrein-related peptidase 5 (KLK5) in human serum

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Open Journal of Clinical Diagnostics 2013年第4期3卷 159-166页

作者： Yanhua Wu Xiaofei Liu Yingjian Chen Chengjin Hu Department of Laboratory Medicine Jinan General Hospital of PLA Jinan China

kallikrein-related peptidases (KLKs) have been proposed as potential cancer biomarkers. kallikrein-related peptidase 5 (KLK5) is a secreted trypsin-like protease of the KLKs. Until now, detection of KLK5 in both biological fluids and tissues has been described frequently due to the potential of being a new cancer biomarker. Our objective was to prepare KLK5 antibodies and establish an ELISA method for KLK5 to study the possible clinical application of KLK5 as a biomarker for malignancies. In this study, recombinant KLK5 protein was produced and purified using a prokaryotic expression system, and then used as immunogen to generate antibodies. High titers of specific antibodies were measured in serum of rabbits after the forth booster injection. And the titer of the antiserum reached 1:106. We have also generated monoclonal antibodies using hybridoma technology and the titer reached 1:105. The activity of KLK5 antibodies was characterized by Western blot and immunohistochemistry. To quantitatively examine KLK5 in serum samples, we established double antibody sandwich ELISA method using mouse mAb as capture and rabbit pAb as tracer antibody. We have detected KLK5 levels in ovarian cancer serum to ensure that our sandwich ELISA measurement to have high sensitivity and specificity. The ranges of linearity reached by the standard curves of the newly developed ELISA were 0.45 ng/mL to 125 ng/mL. The detection limit of the method, defined as the concentration of KLK5 can be distinguished, was 0.20 ng/mL. Median serum KLK5 levels were 3.77 ng/mL and 0.86 ng/mL in ovarian cancer patients and normal female, respectively (P ELISA assay for KLK5. Our preliminary findings prompt that KLK5 may be a new potential biomarker for the diagnosis and prognosis in patients with ovarian.

关键词： Human kallikrein 5 kallikrein-related Peptidases Generation of Antibodies Sandwich ELISA

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Roles of kallikrein-related Peptidase in Epidermal Barrier Function and related Skin Diseases

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International Journal of Dermatology and Venereology 2019年第3期2卷 150-155页

作者： Jiao-Quan Chen Bi-Huang Liang Hua-Ping Li Zi-Yin Mo Hui-Lan Zhu Institute of Dermatology Guangzhou Medical UniversityGuangzhouGuangdong 510095China Department of Dermatology Guangzhou Institute of DermatologyGuangzhouGuangdong 510095China

Introduction The skin barrier usually refers to the physical barrier formed by the cuticle and sebum membrane of the *** barrier not only protects against the onslaught of microorganisms and percutaneous penetration of chemicals and allergens but also prevents the loss of nutrients and moisture from the *** play an important role in maintaining the function of the keraphyllous skin ***,desmocollin,and corneodesmosin are examples of corneodesmosomes.

关键词： kallikrein-related Peptidase Epidermal Barrier Function related Skin Diseases

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