Methods, devices, and reagents for monitoring paclitaxel concentration in plasma for pharmacokinetic-guided dosing of paclitaxel

A pharmacokinetic and paclitaxel technology, which is applied to the determination of paclitaxel in liquid samples and the field of paclitaxel determination, can solve problems such as high cost

Inactive Publication Date: 2017-02-15
AUTOTELIC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the high cost of assays and instrumentation has prohibitive consequences for establishing therapeutic drug monitoring (TDM) for many drugs with relatively narrow therapeutic ranges

Method used

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  • Methods, devices, and reagents for monitoring paclitaxel concentration in plasma for pharmacokinetic-guided dosing of paclitaxel
  • Methods, devices, and reagents for monitoring paclitaxel concentration in plasma for pharmacokinetic-guided dosing of paclitaxel
  • Methods, devices, and reagents for monitoring paclitaxel concentration in plasma for pharmacokinetic-guided dosing of paclitaxel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0187] Determination reagent

[0188] In this example, the preparation of representative detection reagents and capture reagents useful in the assay method and device of the present invention is described.

[0189] Detection reagent: antibody-colloidal gold conjugate . In short, the antibody (see Example 2) was diluted to 1 mg / mL in 0.5X PBS, and the following steps were taken: (1) Shake or vortex the gold to resuspend any precipitated gold, then add 0.5 mL of bare gold The sol is placed in 10 clean individual test tubes; (2) Each tube is marked with a pH value (or 1 to 10) from the provided pH chart; (3) The pH chart is used to add different amounts of buffer to each test tube (4) Place each test tube on a low-speed vortex mixer and add the antibody solution, then mix thoroughly (about 2 to 3 seconds), for 20nm gold, 14μL of 2mg / mL antibody or protein solution is the best; (5) Deep purple and / or black precipitates on some tubes indicate that the antibody or protein is below i...

Embodiment 2

[0194] Paclitaxel antibody

[0195] In this example, the production, processing, purification, characterization and optimization of representative paclitaxel antibodies useful in the methods and devices of the present invention are described.

[0196] Antibody production and processing. The cells were grown in CCM1 (Hyclone) containing 5-10% FBS and 1X Pen / Strep. Once the cell reaches> 1×10 6 The density of cells / mL separates the cells (1:4). The cells were then frozen and stored in 2 separate liquid nitrogen freezing tanks as a backup. Culture the cells in a roller bottle until it reaches 1×10 6 The density of cells / mL. At this time, the culture was no longer fed, and cell viability was monitored daily. Once the cell viability is reduced to <50%, remove the cells and harvest the antibody-rich medium.

[0197] Affinity purification of antibodies. Diafiltration was performed using PBS, pH 7.4, and the harvested antibody was concentrated 10-fold using a 50Kd retention membrane. ...

Embodiment 3

[0256] Representative solid phase competition assay

[0257] In this example, a representative assay demonstrating the efficacy of the solid phase competitive assay is described. This assay demonstrates the usefulness of using the anti-paclitaxel antibody described herein in this type of detection format to provide an informative signal of the presence of paclitaxel in a sample. The results indicate that the changing placement of antibodies can enhance the performance of the assay.

[0258] Paclitaxel lateral flow system. The 1.2 mg / mL BSA-Pac (test line, T) and 0.2 mg / ml goat anti-mouse antibody (control line, C) were striped onto the membrane card (High-Flow Plus HF180 membrane card, Millipore). The anti-paclitaxel antibody-colloidal gold conjugate was absorbed and dried to a binding pad (glass fiber pad, Millipore). In the assay, fetal bovine serum (FBS) spiked with paclitaxel (10 uL) (tracked by 80 μL PBS Tween) was flowed.

[0259] Tandem antibody determination. The antibo...

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Abstract

Methods, devices, and compositions for assaying therapeutic agents. In one aspect, methods, devices, and compositions for assaying paclitaxel to provide therapeutic drug monitoring guided therapy of paclitaxel.

Description

[0001] Cross references to related applications [0002] This application claims the benefits of U.S. Patent Application No. 61 / 975,386 filed on April 4, 2014 and U.S. Patent Application No. 62 / 051,757 filed on September 17, 2014, each of which is expressly incorporated herein by reference in its entirety . [0003] Statement on Sequence Listing [0004] The sequence listing associated with this application is provided in a text format instead of a paper copy, and is hereby incorporated into the specification by reference. The name of the text file containing the sequence listing is 53634_SEQ.txt. The text file is 11KB; it was created on April 6, 2015; and it was submitted with the submission of this manual via EFS-Web. [0005] Background of the invention [0006] Paclitaxel, originally isolated from the bark of the Pacific yew tree, has been established as one of the most effective chemotherapy drugs for a series of cancer types, including lung, ovarian, and breast cancer. The main...

Claims

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Application Information

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IPC IPC(8): G01N33/00G01N33/53
CPCG01N33/558G01N33/94G01N2800/52G01N2407/02G01N33/54388
Inventor 李岳蓉朴哲浩
Owner AUTOTELIC
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