Osteocalcin-N terminal peptide detecting reagent, detection method and application thereof

Abstract

The invention discloses an osteocalcin-N terminal peptide detecting reagent, a detection method and an application thereof. The detecting reagent comprises an immune complex and an antibody detectionsystem. The immune complex uses a first labeled specific antibody, a second labeled specific antibody, an osteocalcin calibration product, an osteocalcin quality control product, and a sample to be tested to compositely form a first labeled specific antibody-osteocalcin-N terminal peptide-second labeled specific antibody. One of the first specific antibody and the second specific antibody is specifically combined with the epitope of an amino acid in an osteocalcin-N terminal peptide amino acid sequence, and the other antibody is specifically combined with the epitope of the amino acid in the osteocalcin-N terminal peptide amino acid sequence. The detecting reagent can specifically detect a completely-carboxylated osteocalcin molecule whole section and a degraded amino terminal N-terminal macromolecule fragment and has the advantages of high sensitivity, good stability and the like. A bone mass loss and a bone conversion rate can be well reflected.

Description

technical field [0001] The invention belongs to the field of reagent diagnosis research, in particular to an osteocalcin-N-terminal peptide detection reagent and its detection method and application. Background technique [0002] The molecular biological characteristics of osteocalcin are as follows: [0003] Osteocalcin (OC) is also known as γ-carboxyglutamic acid-containing protein (BGP), or bone vitamin K dependent protein. Osteocalcin is a non-collagenous protein synthesized and secreted by mature osteoblasts in the bone. It is a component of the bone extracellular matrix, accounting for about 10% to 20% of bone non-collagenous protein. [0004] After translation, human osteocalcin first forms a precursor peptide of 98 amino acid residues, which consists of three parts: the first part is a signal peptide containing 26 amino acid residues, which is detected by receptors on the endoplasmic reticulum membrane Recognize and combine with it, so that the precursor peptide sy...

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Problems solved by technology

[0038] The traditional first-generation osteocalcin assay mainly detects all intact osteocalcin molecules, but the structure of arginine at positions 43-44 of osteocalcin is very unstable, and it degrades rapidly in peripheral blood, making sample pretreatment difficult. As the storage time of the samples prolongs, the detected concentration of osteocalcin decreases; while the second-generation osteocalcin detection is based on the first-generation detection, which detects all intact osteocalcin molecules and degraded molecular fragments. The detected concentration of calcium is stable. Although it solves the problem of easy degradation of osteocalcin molecules, it does not specifically distinguish fully carboxylated and incompletely carboxylated osteocalcin molecules, which leads to the inability to determine the real participation in bone in the blood. The content of transformed osteocalcin is of little significance as a basis for clinical diagnosis

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