Gallium 68 labeled affinity protein PET imaging agent and application thereof
A technology of PET imaging agent and affimer, which is applied in the direction of in vivo radioactive preparations, radioactive carriers, preparations for in vivo experiments, etc., and can solve problems such as insufficient stability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030]Preparation of PDGFR-β Targeted Trimolysin (Z-Tri)
[0031]1.1 Sequence of affinity Z-TRI
[0032]The amino acid sequence of the affinity Z-Tri protein is shown in SEQ ID NO.1.
[0033]1.2 Preparation of the expression plasmid of affinity Z-TRI gene
[0034]The expression plasmid of affinity Z-Tri was constructed by the laboratory pre-design, stored in TOP10 strains (TOP10-PQE30-Z-TRI). Top10-PQE30-Z-Tri hydrochloride was inoculated in a liquid medium containing ampicillin (AMP, 100 μg / ml) LB liquid medium, 37 ° C, 220 rpm shocking overnight, to the bacterial liquid A600To 1. Take some bacterial liquid to extract PQE30-Z-Tri plasmid according to the plasmid extraction kit specification, and the agarose gel electrophoresis is extracted to extract the size of the plasmid, and the company sequencing verifies whether it is PQE30-Z-Tri, which will verify the correct PQE30- The Z-Tri plasmid was stored at 20 ° C.
[0035]1.3 Construction of expression strains Mi5-PQE30-Z-TRI
[0036]The sequence ve...
Embodiment 2
[0054]68Preparation of GA-Z-TRI
[0055]image 3 Shows68A schematic of the GA-Z-Tri affinity probe, where the leftmostmost side is the three polymer probe of the present invention68GA-Z-TRI, its preparation method is as follows:
[0056]3.1Z-TRI and double function chelating agent
[0057]Z-Tri protein 0.5 mg is dissolved in 50 mM sodium hydrogencarbonate solution (pH 8.2 ~ 8.5), with double functional coingent 2 - [(4-isothi cyanophenyl) methyl] -1, 4, 7, 10 - Tetrazo heterocyclic dodecane-1,4,7,10-tetracetic acid (P-SCN-BN-DOTA, referred to as DOTA, after the same) 20 μl (5 moll) was mixed, 37 ° C was incubated at 37 ° C for 2 hours. Using GE PD-10DESALTING Column desalination column, the sodium acetate solution (pH 8.8 ~ 9.0) is a mobile phase, and after removing the unbound DOTA, the successful coupled DOTA-Z-Tri protein is stored in 0.5 m. Sodium acetate (pH 8.8 ~ 9.0) spare.
[0058]3.2Z-Tri68GA tag
[0059]Wash the GE-GA generator using 0.1M super pure hydrochloride68GA CL3The flux, DOTA-Z-T...
Embodiment 3
[0062]68GA-Z-TRI PET imaging effectiveness test
[0063]1) use68GA-Z-Tri bobbin performs PET imaging of subcutaneous humen nude mice to verify its effectiveness.
[0064]The subcutaneous tumor cell line was constructed by subcutaneous tumor cell line, and the malignant glymiloma U-87mg, colon cancer LS-174T, lung cancer A549, osteosarcoma Saos2, breast cancer MDA-MB-231, Hepatic cancer huh7 total 6 malignant tumor skin lower tumor nude mice model.
[0065]After the tumor has a tumor diameter, PET imaging is performed. The specific steps are: 3.7 Mbq of 3.7 Mbq by tail veins from the nude mice after tumor68The Ga-Z-Tri complex protein solution (about 1.5 μg) was developed using MicroPET-CT after 1 hour.Figure 5 As shown, the arrow refers to a tumor, which clearly sees most of the drug through the kidney-bladder pathway to the body, and the corresponding subcutaneous tumor site has obvious radioactive intake, confirmed 68GA-Z-TRI Pixin can be used for tumor PET imaging.
[0066]2) use68Ga-Z-Tri a...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - 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.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap