Abstract
Background: Usually, diphteria toxin concentration and purification are performed after detoxification due to its high danger. In this work, an alternative diphteria toxoid (DTx) concentration and purification method by affinity chromatography is proposed.
Methods: Screening of 19 triazinic dyes for selecting the best ligand for DTx concentration and purification by affinity chromatography was performed.
Results: Cibacron Blue 3GA and Reactive Green 10 showed the highest affinity for DTx. The adsorption isotherms of DTx on Cibacron Blue 3GA-Sepharose and Reactive Green 19-Sepharose showed a good fit of experimental data to a Langmuir-type isotherm and allowed the calculation of a maximum capacity (qm) of 1.21 ± 0.12 µmol DTx/mL matrix and 1.44 ± 0.14 µmol DTx/mL matrix, respectively and a dissociation constant (Kd) of 9.75 ± 2.90 µM and 1.58 ± 0.38 µM, respectively. Blue R-HE-Sepharose isotherm did not show a good fit of experimental data to a Langmuirtype isotherm due to its high Kd.
Conclusion: Cibacron Blue 3GA and Reactive Green 10 are promising ligands for concentration and purification of DTx by affinity chromatography.
Keywords: Diphtheria toxoid, purification, triazinic dyes, affinity chromatography, Cibacron Blue 3GA, ionic exchange chromatography.
Related Books
Indicator Displacement Assays (IDAs): An Innovative Molecular Sensing Approach
Waste Valorization for Value-added Products
Diverse Strategies for Catalytic Reactions
Ultrasound Technology for Fuel Processing
Carbonaceous Quantum Dots: Synthesis And Applications
Recent Advances in Analytical Techniques
Applications of Ionic Liquids in the Oil Industry: Towards A Sustainable Industry
Advances in Combustion Synthesis and Technology
Advanced Techniques of Analytical Chemistry
Applications of Modern Mass Spectrometry
18
2
1
1