Connecting Peptide (C-Peptide) CLIA
52025078
Intended Use
C-peptide (CLIA) is intended for in-vitro quantitative measurement of C-peptide
in human serum or plasma by chemiluminescent immunoassay.
Clinical Significance
C-peptide (C-P) is also called connecting peptide and secreted by the pancreatic 
cells. Proinsulin is the common precursor of C-P and insulin. After enzymatic
digestion, a molecule of proinsulin is cleaved into a molecule of insulin and a
molecule of C-P. C-P is not affected by exogenous insulin. For diabetics who have
been treated with insulin or produced insulin antibodies, the determination of C-
P can evaluate endogenous insulin secretion ability, which is of great significance
for the diagnosis and treatment of diabetes. In addition, C-P is not inactivated by
liver enzymes and has a longer half-life than insulin, instead it is excreted directly
in the urine by the kidneys, so the concentration of C-peptide in the urine within
24 hours can better reflect the function of pancreatic â cells. They are the main
clinical significance of C-peptide detection that the diagnosis of pancreatic
dysfunction case, the analysis of etiology and disease type, and the guidance for
the treatment plan. At present, methods commonly used for C-P detection are
chemiluminescent immunoassay and enzyme-linked immunoassay.
Principle
This C-peptide assay is a “sandwich” chemiluminescent magnetic microparticle
immunoassay. Immunomagnetic microparticles (IMMP), specimen and the
acridinium ester (AE) labelled conjugate are mixed and incubated to form “IMMP-
C-P-conjugate” complexes. After washing, the excess conjugate and other
substances are removed, and then the Chemiluminescence Analyzer Substrate
Buffer is added, the resulting chemiluminescent reaction is quantified in relative
light unit (RLU). The value of RLU is directly proportional to the concentrations of
C-P in the specimen.
Kit Components
Reagent Product Code Description
52025078
C-peptide R1 3.5 mL Magnetic microparticles coated with anti-
human C-peptide monoclonal antibody
C-peptide R2 6 mL Acridinium labelled anti-human C-peptide
monoclonal antibody
C-peptide R3 3.5 mL Phosphate buffer
Calibrator-1 1 mL C-peptide antigen
Calibrator-2 1 mL C-peptide antigen
Plastic Opener 2 nos
* Note: Do not mix or interchange d ifferent batches of kit.
Risk & Safety
Material Safety data sheets (MSDS) will be provided on request
Reagent Storage and Stability
The unopened reagents and calibrators are stable until the expiration dates stated
on the labels, when stored upright at 2-8OC and protected from direct sunlight.
Avoid freezing. After opening, the reagents are stable for 35 days when used on
board, and the calibrators are stable for 35 days at 2-8OC.
Open Vial Stability
Once opened, the reagent casette is stable up to 35 days when kept at 2-8 OC.
On-board Calibration Stability
Calibration is stable for 7 days.
Precaution
Place the reagent kit in upright position always.
For in vitro diagnostic use only.
Package insert must be followed accordingly. Reliability of assay results cannot be
guaranteed if there are any deviations from the instructions in this package insert.
This product requires the handling of human specimens. It is recommended that
all human sourced materials be considered potentially infectious and handled in
accordance with applicable laws. Al l materials contaminated with patient
specimens should be inactivated by validated procedures (autoclaving or chemical
treatment) in accordance with applicable regulations.
 N
www.agappe.com
2 x 50 Test
This product contains chemical ingredients. Contacting with skin or mucosa should
be avoided. If the product is spilled into eyes, mouth or skin accidentally, rinse
with running water and seek for doctor advice if necessary.
This product does not involve human source substances and has no biosafety hazard,
but it should still be considered as a potential infectious substance. The treatment,
use, storage of each component and the disposal of solid and liquid wastes
generated in the analysis process should be hand led in accordance with the
corresponding measures of local biosafety guidelines or regulations.
All test equipment and related glass and plastic wares should be clean and free of
contaminants.
Packaging materials that are non-degradation should be collected and delivered
to the local process center.
Do not use reagent kits beyond the expiration date.
Waste Management
Reagents must be disposed off in accordance with local regulations
Sample
Serum/Plasma
Specimen Col lection&Storage
Follow this package insert as well as the specimen collection tube manufacturer’s
instructions for specimen collection and preparation. Raw materials and addictive
may vary among different manufactures’ collection tubes, which may result in
different results. This kit has not been tested with all collection tube manufactures.
Each laboratory needs to determine the feasibility of the collection tubes or serum
separation products. The specimens should be free of microbial contamination,
fibrin, cells or other particulate matter, severe lipemia, severe hemolysis. If the
sample is cloudy or has visible flocculent, the sample shall be centrifuged, and the
supernatant can be used for detection.
If testing cannot be done before 24 hours after blood collection, the specimens
shall be separated to serum or plasma and then be stored. Specimens may be stored
for up to 24 hours at 2°C~8°C or 3 months at -20°C. Multiple freeze-thaw cycles of
specimens should be avoided. The sample should be transported at low
temperature.
Ensure the patients’ samples, calibrators and controls are at ambient temperature
(15°C~25°C) before measurement.
Do not use mixed specimens.
Volume required: 200 µL for the first c-peptide test plus 50 µL for each additional
c-peptide test from the same sample tube.
Materials provided
C-peptide Casette,Calibrator & Plastic Opener
Procedure
Follow the procedures described in Maccura i1000 Automatic Chemiluminescence
Analyzer User Manual.
Verify that test parameters are loaded before reagents are loaded, for specific
operations, please refer to the analyzer user manual.
Prior to running the kit, ensure that the magnetic microparticles are mixed to keep
them suspended and avoid formation of foam.
Place the reagents in the reagent compartment. When the reagent compartment is
closed, the analyzer will automatically scan and read the reagent RFID information.
Cal ibration
Ensure the calibrator to ambient temperature, mix and place it on the calibrator
rack.
Make sure that the reagent and calibrator lot are same before loading.
Ensure the related calibrator information is obtained by the analyzer, for specific
operations, please refer to the analyzer user manual.
Recommended calibration frequency:
For every new reagent lot.
The controls are out of range.
After 35 days when using the same reagent lot.
After 7 days when using the same reagent kit.
After major repair or replacing spare parts
ISO 9001:2015
EN ISO 13485:2016
Rev.0.280622
 2 x 50 Test
Connecting Peptide (C-Peptide) CLIA 52025078

Quality control Performance

Ensure the control to ambient temperature, mix and place it on the required rack.
Ensure the related control information is obtained by the analyzer for specific
operations, please refer to the analyzer user manual.
Ensure that control values are within the concentration ranges.
Recommended quality control frequency:
For every new reagent.
After every calibration.
Once every 24 hours when the test is in use.
1.Accuracy:
The recovery rate is within the range of 85.0%~115.0%.
2.Sensitivity:
Limit of blank is  0.3 ng/mL The limit of blank is the 95th percentile value of
the zero-concentration calibrator for repeated determination.
Limit of detection is 0.04 ng/mL.
The limit of detection is the lowest analytical concentration which is distinguished
from the limit of blank (Miscalculation probability  = 0.05).

If the quality control procedures in user’s laboratory require more frequent use of Limit of quantification is 0.05 ng/mL.
controls to verify test results, follow the laboratory specific procedures.
3.Linearity:
Limitation
In the range of 0.10ng/mL~30.00ng/mL, the coefficient of correlation r is not less
This assay has only been validated for the specimen type of human serum and
than 0.9900.
plasma (heparin and EDTA). Other types of specimens have not been validated.
4.Precision:
Specimens of severe icterus, hemolysis, lipemia and contamination may result in
error results. The intra-lot coefficient of variation (CV) is not higher than 8.0%.
Interference due to extremely high titers of antibodies to streptavidin and intake The inter-lot coefficient of variation (CV) is not higher than 15.0%.
of high dose of biotin can occur.
5. Interference
Heterophilic antibodies in specimen can react with reagent immunoglobulins,
interfering with in vitro immunoassays. Interfering Substance Concentration (Max)

Patients routinely exposed to animals or animal serum products can be prone to Hemoglobin  5.0 g/L
this interference and anomalous values may be observed. Triglycerides  9.0 mmol/L
The results of different detection systems cannot be used interactively. Bilirubin  400 µmol/L
Reference Range Biotin  20 ng/mL
0.77 ng/mL ~ 3.08 ng/mL RF factor  1200 IU/mL
The above reference interval is the midd le 95% interval of the measurement results
6.HOOK TEST:
of 120 specimens from fasting normal population. It is recommended that each
laboratory establish its own normal range which may be unique to the population There is no high-dose hook effect at C-P concentrations up to 200 ng/mL.
it serves depending upon ethnics, geographical, patient, dietary, or environmental
factors. 7.SPECIFICITY
When the concentration of proinsulin is 10 ng/mL, or the concentration of insulin
Interpretation of Result is 500 µIU/mL, the measured results of C-P are al l not higher than 0.25 ng/mL.
The test results shall be only considered as a clinical reference rather than the BIBLIOGRAPHY
unique basis for confirming or excluding a case. For diagnostic purposes, results 1. Clark PM. Assays for insulin, proinsulin(s) and C-peptide. Ann Clin Biochem,
should always be used in combination with clinical examination, medical history 1999, 36 (5): 541-564.
and other result of inspection.
2. Wahren J, jornval l H.C-peptide makes a comeback. Diabetes Metab Res
Rev,2003,19(5): 345-347.


±


 N

ISO 9001:2015
www.agappe.com EN ISO 13485:2016
Rev.0.280622