Intended use
This immunoassay kit allows for the in vitro quantitative determination of General PYD concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids.
Test principle
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General PYD, During the reaction, General PYD in the sample or standard competes with a fixed amount of biotin-labeled General PYD for sites on a pre-coated Monoclonal antibody specific to General PYD. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General PYD in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Pyridinoline and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	106-116%	108-116%	95-106%	95-105%
EDTA plasma(n=5)	93-103%	108-118%	99-108%	90-100%
heparin plasma(n=5)	101-110%	92-104%	109-119%	96-96%

 

General Annotation

 

Function:

Pyridinoline and deoxypyridinoline were found to be released into the blood during bone degradation and rapidly exereted in the urine. In a preliminary study, both these compounds were proposed as a marker for metastatic bone tumor in patients with prostate cancer.

 

Location:

Pyridinoline is a fluorescent cross-linking compound of collagen fibers. Crosslinks in collagen and elastin are derived from lysyl and hydroxylysyl residues, a process catalyzed by lysyl oxidase. Fujimoto and colleagues first described the isolation and characterization of a fluorescent material in bovine achilles tendon collagen and termed it pyridinoline. It is reported to be present in collagen of bone and cartilage, but is absent in collagen of skin. It is not present in newly synthesized collagen and is formed from aldimine cross-links during maturation of collagen fibers.

 

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