Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay)

DGSX; OCI5; SDYS; SGB; SGBS1; MXR7; Glypican Proteoglycan 3; GTR2-2; Intestinal protein OCI-5; Secreted glypican-3

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 91
EDTA plasma(n=5) 95-102 98
heparin plasma(n=5) 92-99 96
sodium citrate plasma(n=5) 79-93 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 98-105% 96-105% 78-101% 94-101%
EDTA plasma(n=5) 94-105% 83-103% 87-99% 85-104%
heparin plasma(n=5) 93-102% 90-105% 89-96% 85-104%
sodium citrate plasma(n=5) 84-93% 82-89% 83-97% 78-92%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:GPC3) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Nature and Science Diagnostic Role of Serum Glypican-3 as a Tumor Marker for Hepatocellular Carcinoma Sciencepub:10043238
Journal of Applied Pharmaceutical Science Cytotoxic effects of antiglypican-3 against HepG2 cell line Ebscohost: Source
Asian Pacific Journal of Cancer Prevention Can Glypican3 be Diagnostic for Early Hepatocellular Carcinoma among Egyptian Patients? Pubmed: 24460300
Clin Biochem. Diagnostic accuracy of serum glypican-3 for hepatocellular carcinoma: A systematic review and meta-analysis Pubmed: 24362268
Science and Nature Diagnostic Role of Serum Glypican-3 as Hepatocellular Carcinoma Zolcat: Source
Alexandria Journal of Medicine Role of serum glypican-3 in the diagnosis and differentiation of small hepatocellular carcinoma from hepatitis-C virus cirrhosis ScienceDirect: S2090506814000062
J Egypt Natl Canc Inst. Comparing prothrombin induced by vitamin K absence-II (PIVKA-II) with the oncofetal proteins glypican-3, Alpha feto protein and carcinoembryonic antigen in diagnosing hepatocellular carcinoma among Egyptian patients. Pubmed:24841158
European Journal of Pharmacology Evaluation of antiglypican-3 therapy as a promising target for amelioration of hepatic tissue damage in hepatocellular carcinoma Pubmed:25449037
Hepatobiliary Pancreat Dis Int Predictive value of tumor markers in patients with recurrent hepatocellular carcinoma in different vascular invasion pattern abstract:abstract4525.shtml
Clinical and Molecular Hepatology  Glypican-3 level assessed by the enzyme-linked immunosorbent assay is inferior to alpha-fetoprotein level for hepatocellular carcinoma diagnosis pmc:articles
Wiley Glypican‐3: A promising biomarker for hepatocellular carcinoma diagnosis and treatment pubmed:28621802
Biomedicine & Pharmacotherapy Cytotoxic and partial hepatoprotective activity of sodium ascorbate against hepatocellular carcinoma through inhibition of sulfatase-2 and Pubmed:29669302
Journal of Cancer and Clinical Oncology Development and Validation of a Two-Site Immunoradiometric assay for Glypican-3 in plasma: Implication in Diagnosis of Hepatocellular Carcinoma Researchgate:Source
Turkish journal of oncology Serum Glypican-3, Vascular Endothelial Growth Factor, and Interleukin-6 Levels in Hepatocellular Carcinoma 10.5505:tjo.2018.1742
Intensive Care Medicine Experimental Elevated plasma glypicans are associated with organ failure in patients with infection Pubmed: 30618011
VALUE OF SERUM GLYPICAN 3 AS A PREDICTIVE TOOL FOR EARLY DIAGNOSIS OF HEPATOCELLULAR CARCINOMA (HCC)
ADVANCED RESEARCHES Search for Effective Serum Tumor Markers for Early Diagnosis of Hepatocellular Carcinoma Associated with Hepatitis C
Life Sci Selenium nanoparticles overcomes sorafenib resistance in thioacetamide induced hepatocellular carcinoma in rats by modulation of mTOR, NF-κB pathways and … Pubmed:35640776
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