Development of chemiluminescence method based on serum type I collagen hydroxyl terminal peptide β special sequence (β-CTX)

Abstract

The objective of this work is to develop and verify the analytical performance of a chemiluminescence immunoassay for the specific sequence β-carboxy-terminal cross-linking telopeptide of type I collagen (β-CTX) in human serum. Two specific monoclonal antibodies (mAb-8A03 and mAb-3D12) with high affinity for β-CTX were selected, and, under optimized conditions, a chemiluminescence immunoassay method (CLIA) for β-CTX was established. The CLIA of β-CTX detected β-CTX in a wide range of 2.0–6000 ng/L. The recovery rate in serum is 95–105%, the specificity is high, and the cross-reaction rate with common easily interfering substances is low (not more than 0.01%). The CLIA correlates well with Roche electrochemiluminescence immunoassay (ECLIA), with a correlation coefficient of 0.9551, which fully meets the requirements of clinical analysis. The developed β-CTX CLIA kit has high sensitivity and good stability. It has the same performance as the commercial Roche ECLIA kit and can be applied clinically.

Keywords:

• Type I collagen
• type I collagen hydroxyl terminal peptide β specific sequence
• chemiluminescence immunoassay
• osteoporosis
• osteoarthritis
• degenerative bone disease

Author contributions

Song X, and Liu XK conceived and performed the experiment. Wu YF, Huo Z and Shen J provided the samples. Zhang YC, Liang Y, Xu RY and Ma YF screened antibody. Tang XL analyzed the data and wrote the manuscript. All authors have reviewed the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

The study was funded by The National Natural Science Fund of China [82071862, 81872017 and 81572431], The University Natural Science Research Project of Anhui Province [KJ2018ZD011 and KJ2019A0093], The Anhui Provincial Science and Technology Program [202004j07020053 and 1604a0802094], The Research Foundation of the Institute of Environment-Friendly Materials and Occupational Health (Wuhu), Anhui University of Science and Technology [ALW2020YF11] and The Huainan Science and Technology Project [2017B41] funded this research.