Chemiluminiscence kit for detecting human epididymis secretory protein 4

A technology of epididymis secreted protein and chemiluminescence reagent, which is applied in the detection field of human epididymis secreted protein, and can solve problems such as research, HE4-coated antibody and acridinium ester HE4-labeled antibody type research, detection sensitivity can not be optimal, etc.

Active Publication Date: 2020-03-20
普健生物(武汉)科技有限公司
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  • Application Information

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Problems solved by technology

[0005] In the prior art, for example, the patent CN201510240537.X provides a method for the detection of HE4 by acridinium ester chemiluminescence immunology based on gold magnetic particles. The surface of magnetic particles is coupled with HE4-coated antibody, and then the blank site on the surface of gold magnetic particles that is not bound to HE4-coated antibody is blocked, and then the antibody is labeled, combined with the analyte, washed, a

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  • Chemiluminiscence kit for detecting human epididymis secretory protein 4
  • Chemiluminiscence kit for detecting human epididymis secretory protein 4
  • Chemiluminiscence kit for detecting human epididymis secretory protein 4

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Embodiment 1

[0067] In the chemiluminescence kit for detecting human epididymis secretory protein 4 in this example, the selected acridinium ester-labeled 69-G10-B8-G8 detection antibody and biotin-labeled 9-C3-D12-B1 capture antibody are as follows Filter to get:

[0068] S1. Artificially synthesize the human epididymis secretory protein 4 gene, recombine the human epididymis secretory protein 4 gene into the expression vector plasmid pATX1, obtain the HE4-pATX1 expression vector, and verify it by electrophoresis. The results are shown in the appendix figure 1 , by attached figure 1 It can be seen that the cloning site is EcoRI / XhoI, and the human epididymis secretory protein 4 gene gene sequence is shown in SEQ NO.1;

[0069] S2. The HE4-pATX1 expression vector was transfected into 293F cell line and cultured for 6 days, and the supernatant was collected for nickel column purification to obtain human epididymis secretory protein 4; the purified HE4 was subjected to SDS-PAGE electrophore...

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Abstract

The invention discloses a chemiluminiscence kit for detecting human epididymis secretory protein 4. The kit comprises the human epididymis secretory protein 4, an acridinium ester labeled 69-G10-B8-G8detection antibody and a biotin labeled 9-C3-D12-B1 capture antibody, wherein the acridinium ester labeled 69-G10-B8-G8 detection antibody and the biotin labeled 9-C3-D12-B1 capture antibody are respectively obtained by secreting respective corresponding hybridoma cell strains 69-G10-B8-G8 and 9-C3-D12-B1. The two hybridoma cell strains are obtained by performing recombinant plasmid, protein expression, animal immunization and screening and finally subcloning on an artificially synthesized HE4 gene. The double-antibody sandwich compound disclosed by the invention is good in detection sensitivity and precision, wide in linear range, good in antigen reaction specificity and free of cross reaction.

Description

technical field [0001] The invention belongs to the technical field of detection of human epididymis secretory protein, in particular to a chemiluminescent kit for detecting human epididymis secretory protein 4. Background technique [0002] Human epididymis secretory protein 4 (HE4) is a new highly specific ovarian cancer marker, HE4 (also known as WFDC2), mainly composed of WAP-type 4-disulfide-bonded core (WFDC), and extracellular protease inhibitors high homology. The protein family of transcriptional mediators WFDC are protease inhibitors in leukocytes, and HE4 is also postulated to have a role in innate immunity. The protein gene encoded by this gene is located on chromosome 20q12~13.1, with a total length of about 12kb, consisting of 5 exons and 4 introns. The incidence rate of ovarian cancer ranks third among the three major gynecological malignancies. It is not easy to be found because of its inconspicuous early symptoms. With the increase of cancer time, the 5-ye...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/532G01N33/577G01N21/76C07K16/38
CPCG01N33/57449G01N33/532G01N33/577G01N33/57488G01N21/76C07K16/38
Inventor 雷坤代腾飞秦伏波方绪凤鲁亮万定一张永霞
Owner 普健生物(武汉)科技有限公司
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