Related Experiment Video
Updated: Jan 28, 2026
09:10
Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce
Published on: October 18, 2010
13.5K
Simple and rapid analysis of tocilizumab using HPLC-fluorescence detection method
Makoto Takada1, Yoshihito Ohba1, Seitaro Kamiya1
1Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo City, Nagasaki, Japan.
Luminescence : the Journal of Biological and Chemical Luminescence
|February 28, 2019
Summary
A new assay uses HPLC with fluorescence detection to quantify tocilizumab (TCZ) and separate it from Immunoglobulin G subclass 1 (IgG1). This method relies on structural changes induced by organic bases, enabling TCZ determination in blood.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Immunology
Background:
- Tocilizumab (TCZ) is a therapeutic antibody belonging to the Immunoglobulin G subclass 1 (IgG1) class.
- Accurate quantification and differentiation of antibody subclasses in biological samples are crucial for therapeutic monitoring.
- Existing methods may require antigen-antibody reactions, limiting their applicability or efficiency.
Purpose of the Study:
- To develop a novel, rapid, and efficient assay for the determination of tocilizumab (TCZ) in biological matrices.
- To achieve separation of TCZ from other IgG1 antibodies without relying on antigen-antibody interactions.
- To investigate the mechanism underlying the separation of antibody subclasses based on structural modifications.
Main Methods:
- Development of a high-performance liquid chromatography (HPLC) assay coupled with fluorescence detection.
- Implementation of a facile and rapid pretreatment step involving organic solvents and basic compounds (e.g., trimethylamine, triethylamine).
- Analysis of antibody structural changes and their impact on separation characteristics.
Main Results:
- Successful separation of TCZ from IgG1 was achieved using the developed HPLC method.
- The separation mechanism was found to be dependent on the hydrophobicity of the organic bases used in pretreatment, rather than electrostatic charge.
- The pretreatment induced structural changes in antibodies, altering their surface properties and enabling differentiation.
- The assay demonstrated potential for quantifying TCZ in blood samples.
Conclusions:
- A novel HPLC-based assay enables the separation and determination of tocilizumab (TCZ) from IgG1 antibodies.
- The method utilizes organic base-induced structural modifications for antibody subclass differentiation, circumventing the need for antigen-antibody reactions.
- This innovative approach offers a valuable tool for therapeutic drug monitoring of TCZ in clinical settings.
Related Concept Videos
01:17Performing a Simple Data Analysis using MS-Excel Function
1.0K
Microsoft Excel offers a suite of functions and tools ideal for statistical analysis, making it accessible to students and researchers. This article outlines fundamental Excel functions pivotal for data analysis.
SUM: This function calculates the total sum of a range of values. It's the foundation for aggregating data, essential for determining overall trends and totals in datasets.
AVERAGE: It computes the mean value of a given set of numbers, providing a quick insight into the central...
SUM: This function calculates the total sum of a range of values. It's the foundation for aggregating data, essential for determining overall trends and totals in datasets.
AVERAGE: It computes the mean value of a given set of numbers, providing a quick insight into the central...
1.0K
01:18Effects of EDTA on End-Point Detection Methods
660
Different methods, such as visual observance of metal-ion indicators, spectroscopic techniques, and potentiometric methods, can determine the endpoint of an EDTA titration.
In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a...
In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a...
660
01:19Precipitation Titration: Endpoint Detection Methods
6.0K
In argentometric precipitation titrations, endpoints can be detected visually by the Mohr, Volhard, and Fajans methods. In the Mohr method, adding a soluble chromate indicator gives an initial yellow color to the analyte solution. As the titrant is added, the first excess of silver ions forms a red silver chromate precipitate, marking the endpoint. The solution pH should be maintained at about 8 by adding solid CaCO3.
In the Volhard method, a standard excess of AgNO3 is first added to the...
In the Volhard method, a standard excess of AgNO3 is first added to the...
6.0K
01:10Simple Pendulum
8.0K
A simple pendulum consists of a small diameter ball suspended from a string, which has negligible mass but is strong enough to not stretch. In our daily life, pendulums have many uses, such as in clocks, on a swing set, and on a sinker on a fishing line.
The period of a simple pendulum depends on two factors: its length and the acceleration due to gravity. The period is completely independent of any other factors, such as mass or maximum displacement. For small displacements, a pendulum is...
The period of a simple pendulum depends on two factors: its length and the acceleration due to gravity. The period is completely independent of any other factors, such as mass or maximum displacement. For small displacements, a pendulum is...
8.0K
01:21Simple Trusses
2.9K
A truss is a structural framework consisting of slender members connected at joints, designed to support external loads while minimizing material usage and weight. Simple trusses are a type of planar truss where all members lie within a single two-dimensional plane.
The most basic planar truss is a simple truss with three members arranged in a triangular formation. This triangular truss is inherently stable and rigid due to its geometry, making it an ideal starting point for creating more...
The most basic planar truss is a simple truss with three members arranged in a triangular formation. This triangular truss is inherently stable and rigid due to its geometry, making it an ideal starting point for creating more...
2.9K
01:21Simple Harmonic Motion
15.1K
Simple harmonic motion is the name given to oscillatory motion for a system where the net force can be described by Hooke's law. If the net force can be described by Hooke's law and there is no damping (by friction or other non-conservative forces), then a simple harmonic oscillator will oscillate with equal displacement on either side of the equilibrium position. To derive an equation for period and frequency, the equation of motion is used. The period of a simple harmonic oscillator is given...
15.1K