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TRPM8 protein and related polypeptide fragment and new application of antibody thereof

A polypeptide fragment and protein technology, applied in the treatment of chronic prostatitis/chronic pelvic pain syndrome, in the field of monoclonal or polyclonal antibodies, can solve the problem of unclear pathophysiological changes, huge patient population, and difficult diagnosis and treatment of prostatitis. And other issues

Inactive Publication Date: 2016-02-10
韩蕾
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, its huge patient population and high medical expenses have caused a large economic burden on public health.
[0004] Since the etiology and pathogenesis of chronic prostatitis or chronic pelvic pain syndrome have not yet been clarified, and the pathophysiological changes are still not very clear, there are many problems in the diagnosis of prostatitis, the judgment of the severity of the disease, the choice of treatment methods, and the evaluation of curative effect. On the one hand, there are no clear standards to follow at home and abroad, so most clinicians currently find it difficult to diagnose and treat prostatitis clinically.

Method used

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  • TRPM8 protein and related polypeptide fragment and new application of antibody thereof
  • TRPM8 protein and related polypeptide fragment and new application of antibody thereof
  • TRPM8 protein and related polypeptide fragment and new application of antibody thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Preparation of TRPM8-related polypeptide fragments

[0047] From the transmembrane analysis of TRPM8: there are the following sections outside the membrane: 1-692, 758-796, 849-862, 916-953, 1025-1104. Based on various software analysis, the following six extracellular segments of transmembrane proteins were finally selected, and the selected amino acid sequences basically covered the extracellular segments. which is:

[0048] T-1: KINTKANDTSEEMRHRFRQLDTKLND(26AA);

[0049] T-2: FKNEDNETLAWEGVMKENYL (20AA);

[0050] T-3:DGTTYDFAHCTFTGNESKPL(20AA);

[0051] T-4: VSRNLGPKIIMLQ(13AA);

[0052] T-5:DEVRQWYVNGVNYFTD(16AA);

[0053] T-6: LTVIKMEEAGDEIVSNA (17AA).

[0054] The above peptides were synthesized by entrusting Wuhan Baiyixin Biotechnology Co., Ltd. TRPM8 protein was purchased from ProteinTechGroup.

Embodiment 2

[0055] Example 2 Detection of TRPM8 protein and TRPM8-related polypeptide fragment antigenicity

[0056] In order to prove that TRPM8 protein is the key to the induction of CP / CPPS and the antigenic pathogenicity of TRPM8-related polypeptide fragments, the following experiments were carried out. ) to establish an animal model of CP / CPPS, and compared it with the prostatic protein homogenate modeling method. At the same time, the TRPM8 in the prostatic protein homogenate was removed by using the immunomagnetic bead method, and it was used as a negative control for modeling. The HE staining method was used to evaluate the severity of inflammation in the prostate tissue of the animals in each model group, and the severity of the inflammation in the prostate was scored pathologically; the CD3 immunofluorescence method was used to observe the infiltration of T lymphocytes in the prostate of the model animals .

[0057] Forty 8-week-old SPF-grade male SD rats, weighing 250±20 g (p...

Embodiment 3

[0076] Example 3 Preparation of TRPM8 protein and TRPM8-related polypeptide fragment antibodies

[0077] (1) Selection and immunization of animals

[0078] 1. Selection of animals: purebred BALB / C mice.

[0079] 2. Immunization scheme: Selecting an appropriate immunization scheme is crucial to the success of cell fusion hybridization and obtaining high-quality McAb. Generally, the first immunization is started according to the established immunization scheme about two months before the fusion, and the immunization scheme should be determined according to the characteristics of the antigen.

[0080] The full sequence of human TRPM8 protein and the polypeptide fragments shown in SEQ ID No: 1-6 were selected as antigens, and mice were immunized with complete Freund's adjuvant.

[0081] The initial immunization antigen 1 ~ 50μg plus Freund's complete adjuvant multi-point subcutaneous injection or intrasplenic injection (generally 0.8 ~ 1ml, 0.2ml / point). After 3 weeks, the se...

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Abstract

The invention discloses a TRPM8 related polypeptide fragment. Amino acid sequences are shown in the SEQ ID No:1-6. The invention further discloses a TRPM8 protein, the TRPM8 related polypeptide fragment and application of the corresponding antibody of the TRPM8 related polypeptide fragment for preparing a chronic prostatitis or a chronic pelvic pain syndrome diagnostic reagent. A TRPM8 protein molecule or a TRPM8 related polypeptide fragment or the antibody level in liquid of a patient is detected, the chronic prostatitis or chronic pelvis pain comprehensive syndrome is effectively diagnosed, and other prostate diseases can be effectively distinguished. In addition, monoclonal or polyclonal antibodies of 1000-3000 IU TRPM8 protein or related polypeptide fragments are injected through vein or under the skin, and clinical features of the chronic prostatitis or the chronic pelvis pain syndrome can be cured or obviously relieved.

Description

technical field [0001] The invention belongs to the field of immunodiagnostic reagents and the field of therapeutic drugs, and specifically relates to the application of a TRPM8 protein, related polypeptide fragments or antibodies thereof in the diagnosis of chronic prostatitis / chronic pelvic pain syndrome and monoclonal or polyclonal antibodies against the protein Use in the treatment of chronic prostatitis / chronic pelvic pain syndrome. Background technique [0002] Prostatitis is a common disease in adult men. The incidence rate of prostatitis symptoms in male population in my country is 8.4%, and the outpatients account for 8-25% of the total outpatients in urology department. Statistics show that about 50% of men will be affected by prostatitis at some point in their lives. [0003] Prostatitis is currently divided into four types, of which type Ⅲ, namely: chronic prostatitis (Ⅲa type) / chronic pelvic pain syndrome (Ⅲb type) (chronic prostatitis / chronic pelvic pain sy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47G01N33/68G01N33/577G01N33/558G01N33/543G01N33/535A61K39/395A61P13/08A61P15/00
CPCC07K14/4713A61K2039/505C07K16/18G01N33/535G01N33/54306G01N33/558G01N33/577G01N33/6893
Inventor 韩蕾周晓辉周雨晏周莉
Owner 韩蕾
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