Very Helpful HB-EGF ELISA Kit for Human Samples

September 10, 2018

Mercy Health
Laboratory
Student Researcher

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Company:

R&D Systems

Product Name:

HB-EGF ELISA

Catalog Number:

DY259B

Background: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a rational target for ovarian cancer therapy. The aim of this study was to examine HB-EGF levels in the peritoneal fluid and serum of ovarian cancer (OVCA) patients. Patients and Methods: Samples were collected from six healthy women, 21 OVCA patients, and 21 ovarian cyst patients. HB-EGF levels were measured using a sandwich ELISA kit and calculated using a parallel line assay. Results: No significant difference between the slopes of the standard and sample curves was observed at an anti-HB-EGF antibody concentration of 1.6 μg/ml. HB-EGF levels in the peritoneal fluid and serum of OVCA patients were significantly higher than those in patients with ovarian cysts or controls. Serum HB-EGF levels were also significantly correlated with levels in peritoneal fluid in OVCA patients. Conclusion: We developed an assay for the exact measurement of HB-EGF levels in peritoneal fluid and serum.

Experimental Design and Results Summary

Application

ELISA

Starting Material

Human peritoneal fluid and serum

Protocol Overview

A 96-well plate was coated with 100 μl of SCA at concentrations of 0.4, 0.8, 1.6 or 3.2 μg/ml according to the modified manufacturer's protocol. The slopes of the standard and sample curves at each concentration of SCA were estimated and statistically compared using analysis of covariance (ANCOVA). When no significant difference was observed between the standard and sample slopes, the concentration of HB-EGF in the peritoneal fluid or serum was calculated using a parallel line assay method.

Tips

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Results Summary

Relationship between HB-EGF levels in peritoneal fluid and in serum in OVCA patients. Six sets of peritoneal fluid and serum samples were obtained from the same OVCA patient. The correlation between HB-EGF levels in the peritoneal fluid and serum was analyzed to evaluate the clinical significance of serum HB-EGF levels in OVCA. The results showed a significant correlation between HB-EGF levels in peritoneal fluid and serum

DOI or PMID #

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Additional Notes

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Image Gallery

Summary

The Good

To accurately measure HB-EGF levels in the peritoneal fluid and serum of OVCA patients, a higher concentration of SCA was needed, suggesting the possibility that many unknown molecules may bind to sHB-EGF in patients with OVCA, which may reduce the affinity of HB-EGF for the SCA.

The Bad

HB-EGF levels in (A) serum and (B) peritoneal fluid of patients with ovarian cancer (OVCA), ovarian cysts, and healthy controls. Horizontal lines indicate the mean values. not significant

The Bottom Line

The levels of HB-EGF in the peritoneal fluid and serum of OVCA patients was markedly increased, and a positive correlation between HB-EGF levels in peritoneal fluid and serum was observed for OVCA patients. This suggests that the expression level of HB-EGF in the serum directly reflects the amount of HB-EGF that cancer cells secrete into the peritoneal cavity. In conclusion, the results of the present study show that we developed an accurate method for HB-EGF determination in peritoneal fluid and serum samples.

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