Tacrolimus monoclonal antibody hybridoma cell strain and application thereof
A hybridoma cell line and monoclonal antibody technology, which is applied in the field of immunoassay, can solve the problems of cumbersome steps, rapid detection, and high cost, and achieve good detection sensitivity and specificity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0022] Example 1 Preparation of hybridoma cell line ICP2B3
[0023] (1) Preparation of complete antigen:
[0024] a. The synthetic route of hapten is as follows:
[0025]
[0026] Weigh 19 mg of tacrolimus, dissolve it in 0.6 mL of methanol:water:pyridine (volume ratio 4:1:1), add 13 mg of carboxymethoxylaminehemihydrochloride (CMO), and stir in a water bath at 70°C 6 h, and stand at room temperature overnight in the dark. Subsequently, the solution was dried by nitrogen gas, resuspended in 1 mL of chloroform, and extracted three times with ultrapure water. The organic phase was collected and dried again with nitrogen gas, and the final product was dissolved in 1 mL of anhydrous N,N-dimethylformamide (DMF), namely the tacrolimus hapten.
[0027] b. Weigh 10 mg of tacrolimus hapten and dissolve it in 800 μL DMF, then add 6.3 mg 1-ethylcarbodiimide hydrochloride (EDC), 5 mg N-hydroxysuccinimide (NHS), the mixture was stirred at room temperature for 6 h; 5 mg keyhole limpe...
Embodiment 2
[0037] Example 2 IC of tacrolimus monoclonal antibody 50 Determination of
[0038] Carbonate buffer solution (CBS): Weigh out Na 2 CO 3 1.59g, NaHCO 3 2.93g, respectively dissolved in a small amount of double distilled water and mixed, add double distilled water to about 800mL and mix evenly, adjust the pH value to 9.6, add double distilled water to make up to 1000 mL, store at 4°C for later use;
[0039] Phosphate buffered saline (PBS): 8.0 g NaCl, 0.2 g KCl, 0.2 g KH 2 PO 4 , 2.9 g Na 2 HPO 4 • 12 H 2 O, dissolve in 800mL pure water, adjust the pH to 7.2-7.4 with NaOH or HCl, and set the volume to 1000mL;
[0040] Washing solution (PBST): add 0.5 mL of Tween-20 to 1000 mL of 0.01 mol / L PBS solution with pH 7.4;
[0041] PBST: PBS containing 0.05% Tween-20;
[0042] Antibody diluent: wash buffer containing 0.1% gelatin;
[0043] TMB Chromogenic Solution: Solution A: Na 2 HPO 4 •12H 2 O 18.43 g, citric acid 9.33 g, dilute to 1000 mL with pure water; solution B:...
PUM
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