The development and characterization of an ELISA specifically detecting the active form of cathepsin K

TY - JOUR

T1 - The development and characterization of an ELISA specifically detecting the active form of cathepsin K

AU - Sun, S

AU - Karsdal, M A

AU - Bay-Jensen, A C

AU - Sørensen, M G

AU - Zheng, Q

AU - Dziegiel, Morten Hanefeld

AU - Maksymowych, W P

AU - Henriksen, K

N1 - Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

PY - 2013/10

Y1 - 2013/10

N2 - OBJECTIVE: Cathepsin K plays essential roles in bone resorption and is intensely investigated as a therapeutic target for the treatment of osteoporosis. Hence an assessment of the active form of cathepsin K may provide important biological information in metabolic bone diseases, such as osteoporosis or ankylosing spondylitis. METHODS: Presently there are no robust assays for the assessment of active cathepsin K in serum, and therefore an ELISA specifically detecting the N-terminal of the active form of cathepsin K was developed. RESULTS: The assay was technically robust, with a lowest limit of detection (LOD) of 0.085ng/mL. The average intra- and inter-assay CV% were 6.60% and 8.56% respectively. The dilution recovery and spike recovery tests in human serum were within 100±20% within the range of the assay. A comparison of latent and active cathepsin K confirmed specificity towards the active form. Quantification of the levels of active cathepsin K in supernatants of purified human osteoclasts compared to corresponding macrophages showed a 30-fold induction (p

AB - OBJECTIVE: Cathepsin K plays essential roles in bone resorption and is intensely investigated as a therapeutic target for the treatment of osteoporosis. Hence an assessment of the active form of cathepsin K may provide important biological information in metabolic bone diseases, such as osteoporosis or ankylosing spondylitis. METHODS: Presently there are no robust assays for the assessment of active cathepsin K in serum, and therefore an ELISA specifically detecting the N-terminal of the active form of cathepsin K was developed. RESULTS: The assay was technically robust, with a lowest limit of detection (LOD) of 0.085ng/mL. The average intra- and inter-assay CV% were 6.60% and 8.56% respectively. The dilution recovery and spike recovery tests in human serum were within 100±20% within the range of the assay. A comparison of latent and active cathepsin K confirmed specificity towards the active form. Quantification of the levels of active cathepsin K in supernatants of purified human osteoclasts compared to corresponding macrophages showed a 30-fold induction (p

U2 - 10.1016/j.clinbiochem.2013.04.012

DO - 10.1016/j.clinbiochem.2013.04.012

M3 - Journal article

C2 - 23623829

SN - 1873-2933

JO - Clinical Biochemistry

JF - Clinical Biochemistry

ER -