Diagnosis and treatment of cancer using anti-GPR49 antibody
An antibody and cancer technology, applied in the direction of antibodies, antibody mimetics/scaffolds, preparations for in vivo experiments, etc., can solve the problems such as hyperexpression of which are not seen
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0265] (1) Preparation of effector cells
[0266] Spleens are removed from CBA / N mice and the like, and spleen cells are isolated in RPMI1640 medium (manufactured by Invitrogen). Wash with the same medium containing 10% fetal bovine serum (FBS, produced by HyClone), and then adjust the cell concentration to 5 × 10 6 cells / mL to prepare effector cells.
[0267] (2) Preparation of complement solution
[0268] A complement solution can be prepared by diluting Baby Rabbit Complement (manufactured by CEDARLANE) 10 times with a medium containing 10% FBS (manufactured by Invitrogen).
[0269] (3) Preparation of target cells
[0270] Cells expressing GPR49 protein were treated with 0.2mCi of 51 Cr-sodium chromate (manufactured by GE Healthcare Bio-Sciences) was incubated together in DMEM medium containing 10% FBS at 37° C. for 1 hour to radiolabel the target cells. As cells expressing GPR49 protein, cells transformed with a gene encoding GPR49 protein, gastric cancer, colon cance...
Embodiment 1
[0388] [Example 1] GPR49 mRNA expression analysis using human exon 1.0ST chip
[0389] In order to clarify the expression distribution of human GPR49 mRNA in clinical cancers, cancer cell lines, and various normal organs, expression analysis was performed using the human exon 1.0ST chip (Affymetrix) originally developed for splice variant analysis. The advantage of using the human exon 1.0ST chip for expression analysis is that compared with the existing expression chip of Affymetrix, which basically has only one probe set on the 3' side of each gene, the human exon 1.0ST At least one probe set can be set for each exon of the gene in the chip, so when using this chip to perform expression analysis on each gene, the expression data of multiple probe sets can be obtained for each gene. The reliability of expression data has been improved.
[0390] In this expression analysis, tumor sections from excised tissues of 22 lung adenocarcinomas, 13 tumors of gastric cancer, 5 tumors...
Embodiment 2
[0396] [Example 2] Establishment of cells expressing full-length human GPR49
[0397] Based on NCBI retrieval numbers NP_003658.1 (SEQ ID NO: 1 (amino acid sequence)) and NM_003667.2 (SEQ ID NO: 2 (nucleotide sequence)), the full-length human GPR49 cDNA was isolated by PCR, and This was cloned into a mammalian cell expression vector (pcDNA5 / FRT / TO) (Invitrogen). pcDNA5 / FRT / TO is a hybrid human CMV / TetO 2 The gene inserted under the promoter can be induced to express and is integrated with a neomycin resistance gene as a vector for drug resistance markers. Furthermore, the full-length human GPR49 cDNA was introduced into 293FlpIn T-Rex cells using the expression system FlpIn System (Invitrogen), which can induce expression only in the presence of tetracycline or doxycycline. In DMEM (high sugar) / 10%FBS / 100μg / mL Zeocin TM (Invitrogen) / 15 μg / ml blasticidin (Invitrogen) to introduce the expression vector into 293FlpIn T-Rex cells cultured with Fugene6 (Roche). According to t...
PUM
Property | Measurement | Unit |
---|---|---|
molecular weight | aaaaa | aaaaa |
molecular weight | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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