Development and clinical application of a Sandwich ELISA for hK6 detection
10.3969/j.issn.1673-4130.2017.17.001
- VernacularTitle:人组织激肽释放酶6双抗夹心ELISA方法的建立及临床初步应用
- Author:
Hua ZHANG
;
Chengjin HU
;
Kuixiang CHEN
- Keywords:
human kallikrein 6;
monoclonal antibody;
enzyme-linked immunosorbent assay
- From:
International Journal of Laboratory Medicine
2017;38(17):2337-2338,2341
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare the mAbs against hK6 for establishing a sandwich enzyme-linked immunosorbent assay(ELISA) of hK6,and exploring its clinical value.Methods The hybridoma technique was used to prepare mAbs against hK6.The mAbs were purified and labelled with horseradish peroxidase for the sandwich ELISA method.The sandwich ELISA method was used to detect the serum hK6 concentrations in patients with malignant gastric neoplasm.Then the best antibody pair was selected from coating antibody and enzyme-linked antibody to establish a sandwich ELISA method through the chessboard titrations.Compared with CEA,we explored the feasibility of hK6 as gastric cancer biomarkers.Results A sandwich ELISA method was established for quantifying hK6 in serum.The results showed that the optimal concentration of coating antibody was 5 μg/mL.The optimal concentration of enzyme-linked antibody was 1:2 000.Serum hK6 in the patients with gastric cancer groups[(5.78±1.66)ng/mL] than healthy individuals groups[(3.35±0.67)ng/mL] and those in gastric ulcer groups[(3.59±1.02)ng/mL],the difference was statistically significant(P<0.05).Furthermore,there was no significant difference in values of serum hK6 between patients with gastric ulcer groups and healthy individuals groups(P>0.05).The hK6 positive rate of gastric cancer was 69.70％,and CEA was 45.46％.In the combined detection,the positive rate was 78.79％.Conclusion A sandwich ELISA is established successfully.As a favorable serum biomarker for gastric cancer,the detection of hK6 together with CEA is helpful in the diagnosis of gastric cancer.
