Adaptation of the pore diffusion model to describe multi-addition batch uptake high-throughput screening experiments

Area of Science:

• Biophysical Chemistry
• Chemical Engineering
• Chromatography

Background:

• Equilibrium isotherm and kinetic mass transfer measurements are crucial for chromatographic column modeling.
• Traditional methods are time-consuming and labor-intensive, limiting high-throughput applications.
• Existing high-throughput methods face constraints in protein concentration, incubation, and resin volumes.

Purpose of the Study:

• To develop a novel, time-efficient technique for measuring protein uptake kinetics in a high-throughput format.
• To address the limitations of current methods in protein addition, incubation, and separation times.
• To adapt and validate the pore diffusion model for multi-addition batch uptake measurements.

Main Methods:

• Developed a multi-addition batch uptake technique using sequential protein additions and separation in a 96-well plate format.
• Adapted the pore diffusion model to simulate multi-addition batch uptake kinetics.
• Compared the novel technique with traditional batch uptake measurements for an Fc-fusion protein on an anion exchange resin.

Main Results:

• The multi-addition batch uptake technique demonstrated acceptable agreement with traditional methods under tested conditions.
• The adapted pore diffusion model effectively modeled the multi-addition batch uptake process.
• A sensitivity analysis of the model's physical inputs was performed.

Conclusions:

• The multi-addition batch uptake technique offers a viable and efficient alternative for high-throughput kinetic measurements.
• This method enhances the speed and practicality of characterizing protein-resin interactions.
• Further exploration of the technique's advantages and limitations is warranted.