Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

LRP16 monoclonal antibody hybrid oncocyte strain and uses thereof

Inactive Publication Date: 2008-03-12
GENERAL HOSPITAL OF PLA
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] A large number of studies have shown that ERα is closely related to the treatment and prognosis of estrogen-dependent tumors, but clinicians also realize that the detection of ERα expression in tumors by biochemical and immunohistochemical methods cannot accurately reflect the ERα signaling pathway This is an important cause of resistance to endocrine therapy in breast cancer and an important limiting factor in the selection of tumor chemoradiotherapy strategies

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • LRP16 monoclonal antibody hybrid oncocyte strain and uses thereof
  • LRP16 monoclonal antibody hybrid oncocyte strain and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Preparation of hybridoma cells

[0024] 1. Materials and sources

[0025] 1. Antigen: LRP16 protein (GeneBank No.AF202922), according to the literature (Huang Ke, Meng Yuanguang, Han Weidong, Zhao Yali, Li Qi, Song Lei. Exploration of preparation and expression of polyclonal antibody of LRP16 gene. Journal of Military Medical Training College. 2007, 28 (2): 87-89.) prepared by the method. (LRP16 protein was dissolved in 1×PBS, adjusted concentration: 4mg / ml)

[0026] 2. Animals: ABLB / c mice aged 6-8 weeks (Experimental Animal Center of the Academy of Military Medical Sciences of the Chinese People's Liberation Army)

[0027] 3. Myeloma cell line: SP2 / 0 (US ATCC cell bank)

[0028] 4. Complete and incomplete adjuvant (SIGMA company)

[0029] 5. RPMI-1640 basal medium, incomplete RPMI-1640 medium, complete RPMI-1640 medium, HT medium, HAT medium (Gibico company)

[0030] 2. Method

[0031] 1. Immunization of animals:

[0032] Using stirring and mixing...

Embodiment 2

[0046] Embodiment 2: ELISA method screening hybridoma cells

[0047] 1. Main reagents:

[0048] 1. Coating solution: carbonate buffer pH=9.6, Na 2 CO 3 1.59g; NaHCO 3 2.93g; add distilled water to 1000ml.

[0049] 2. Rinse and diluent: PBST pH=7.4: NaCl 8.0g; KH 2 PO 4 0.2g; Na 2 HPO 4 .12H 2 O2.9g; KCl 0.2g; Tween 0.5ml add distilled water to 1000ml.

[0050] 3. Blocking solution: 1% BSA (1g / 100ml BSA purchased from SIGMA Company)

[0051] 4. Chromogenic solution: TMB, add 3ul 2mg / ml antioxidant Na per ml TMB 2 S2O 3 .5H 2 o

[0052] Liquid A: 0.2M Na 2 HPO 4 (28.4g / L) 25.7ml

[0053] 0.1M citric acid (19.2 g / L) 24.3ml

[0054] h 2 o 2 0.1%; Add 50ml of distilled water.

[0055] Liquid B: TMB 3.90g

[0056] Citric acid 10.52g

[0057] EDTA 1.86g

[0058] Glycerin 2000ml

[0059] DMSO 300ml, heat to dissolve and add distilled water to 10000ml.

[0060] 5. Stop solution (2M H 2 SO 4 ): 178.3ml of distilled water, 21.7ml of concentrated sulfuric acid ...

Embodiment 3

[0077] Example 3: Expression of monoclonal antibodies in animals

[0078] 1. Materials and sources

[0079] 1. Mice: adult BALB / c mice, aged >15 weeks, (Experimental Animal Center, Academy of Military Medical Sciences, Chinese People's Liberation Army)

[0080] The body weight before abdominal treatment was 30.0g for size 1 and 29.1g for size 2

[0081] The body weight after treatment with liquid paraffin is 31.2g, No. 2 30.6g

[0082] 2. Cell line: CGMCC No.1992.

[0083] 3. Culture medium selection: 8%-9% fetal bovine serum complete medium, (Germany Biochrom AG company)

[0084] 2. Method

[0085] 1. Select SP2 / 0 cells in a good growth state for expansion and culture. The cell growth density should not be too large, preferably not exceeding 80% of the bottom area. About each mouse needs 9-12 flasks (25cm culture flask) of cells, and each flask contains about 5×10 6 cell.

[0086] 2. Intraperitoneal treatment: intraperitoneally inject 0.7ml of high-pressure liquid paraff...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a LRP16 single cloned antibody hybrid tumor cell line CGMCC 1992 and the application of the cell line, and pertains to the technical field of medicine biology. The advantages with the invention are: 1) the chromosome is steady, and the cell line will secrete IgG1 antibody; 2) the cell line can be cultured in a no-serum culturing system; 3) the titer of the secreted antibody is high, the upper serum directly cultured can be up to 1:200,000; 4) abdominal dropsy of rat can be up to 1 mg / ml; the titer can be up to 1: 3,600,000, and can be purified by protein G; 5) the secreted antibody can be used for immunity combination (1:1000) and immunity mark (western blot) test (1:3000).

Description

technical field [0001] The invention relates to an LRP16 monoclonal antibody hybridoma cell line and an application thereof, belonging to the field of medical biotechnology. Background technique [0002] Hormone-related tumors, mainly including breast cancer, endometrial cancer and ovarian cancer, are the most common malignant tumors in women. Estrogen overstimulation and estrogen receptor α (ERα) overexpression are key factors in the occurrence and progression of tumors. The results of the study show that accurately judging the functional state of ERα in tumor cells is an important indicator for prognosis and selection of treatment strategies. For example, patients with ERα-negative breast cancer have better response to chemotherapy than patients with positive breast cancer, and their response to endocrine therapy is better than that of patients with ERα-positive breast cancer. Poor, ERα-positive breast cancer patients can be divided into lymph node metastasis group and no ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/18C07K16/18G01N33/577G01N33/74
Inventor 孟元光韩为东陈美霞张燕赵亚力伍志强
Owner GENERAL HOSPITAL OF PLA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com