Abstract
Serum P1NP, one of the important biomarkers for bone turnover, is commonly used for the prediction of bone fracture and the prognosis of osteoporosis after therapy. We developed a P1NP chemiluminescence assay and evaluated changes in bone metabolism markers in lung transplant patients. The screened 2 P1NP antibodies with constructed antigens and α-1 chain antigens expressed by the Corynebacterium glutamate expression system were applied into assay development. The assay performance was evaluated to examine the reliability. A normal Q-Q plot was used to establish male reference interval. Changes of bone metabolism markers before and after lung transplantation in 19 patients were evaluated. The linear factor R of P1NP reagent was greater than 0.99. The limit of detection was 3.32 ng/ml. The precision of the three batches of P1NP reagents was lower than 8%. Method comparison with Roche P1NP reagent showed that the correlation coefficient R2 was 0.91. In the monitoring of bone mass in a short time, bone metabolism markers can better indicate the change of bone mass, while the traditional bone mineral density detection is lagging behind the bone metabolism markers. P1NP and β-CrossLap to bone mass change in patients after lung transplantation, and P1NP and β-CrossLap are very good clinical markers for bone mass monitoring.
Highlights
Serum P1NP, introduced by the IOF-IFCC Bone Marker Standards Working Group, is one of the key biomarkers indicating bone formation
Bone turnover markers (BTMs) are parameters that were widely measured in metabolic bone diseases, including osteoporosis, to evaluate the severity of the disease
Dynabeads MyOne streptavidin-precoated beads, EZLink Sulfo-NHS-LC-Biotinylation Kit, succinimidyl 4-(N-maleimidomethyl) cyclohexane-1carboxylate, and 4′-hydroxyazobenzene-2-carboxylic acid (HABA) solution were obtained from ermo Fisher (Waltham, MA, USA); perfluorohexanoate, methanol, EDC, and NHS; horseradish peroxidase (HRP) was purchased from BBI Solutions (Portland, ME, USA); streptavidin was purchased from Hangzhou NeuroPeptide Biological Science and Technology Incorporation, Ltd. (China). e AKTA purifier system was purchased from GE Healthcare (Chicago, IL, USA); microscopes were purchased from Olympus (Tokyo, Japan); and an automagnetic beads chemiluminescent analyser was supplied by Baiming Biotechnology (Yuncheng, China)
Summary
Serum P1NP, introduced by the IOF-IFCC Bone Marker Standards Working Group, is one of the key biomarkers indicating bone formation. Human serum or plasma P1NP level was indicative for either the prediction of bone fracture or the monitoring of prognosis of osteoporosis patients after receiving therapies. Osteoporosis is one of the most prevalent metabolic bone diseases and is especially prevalent among elderly people worldwide. Low bone mass, measured as bone mineral density (BMD), is often associated with increased risk of bone fracture and is the leading cause of morbidity and mortality of osteoporosis [1]. Bone turnover markers (BTMs) are parameters that were widely measured in metabolic bone diseases, including osteoporosis, to evaluate the severity of the disease. Because of the significance in studying the bone health, numerous assays were developed and are currently available [2]
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