CEInfinite: icIEF Fractionation
Fractionate all the material you need in a fraction of the time.
Unattended, reproducible collection of icIEF peaks for deeper molecular characterization.
Late-stage biopharmaceutical development often necessitates the in-depth interrogation of protein charge isomers to understand their impact on drug stability and efficacy.
While icIEF is a popular tool for characterizing charge heterogeneity, it traditionally requires the parallel use of ion exchange chromatography (IEX) to fractionate charge variants for further analysis. This process can be time-consuming, and the IEX separation profile is not always comparable to icIEF.
With the introduction of icIEF fractionation on the CEInfinite, you can directly probe icIEF peaks to build confidence in your molecule and get it over the finish line faster. CEInfinite enables unattended, reproducible fractionation for the collection of icIEF peaks in quantities suitable for in-depth peptide mapping, amino acid sequencing, potency assays, and more.
CEInfinite icIEF Fractionation Workflow
Step 1: Focusing (15 min)
Inject the sample into the icIEF cartridge to separate and focus charge isomers at 3000 V.
Step 2: Mobilization (30 min)
Initiate the pressure mobilization and makeup flow to drive the icIEF peaks toward the collection vials.
Step 3: Fraction Selection and Verification (60 min)
Define each fraction interval during Step 2 and then confirm their purity by analytical icIEF
Step 4: Automatic Fractionation (45 min/run)
Use the settings from Steps 1-3 to automate the fraction collection process for 20-30 runs per day.
Why Choose icIEF?
- High Reproducibility: AES’ patented technology ensures high reproducibility
- High Resolution & High Purity: the peaks with ≤0.1 pI difference can be fractionated with high purity up to 99%
- High Yield: collect as much as 40 µg/run and 35 runs/day, each run ≤ 1 hour
- Less Labour: automated, temperature-controlled, and minimal user intervention
Applications for icIEF Fractionation on CEInfinite
Create new opportunities and safeguards during biopharmaceutical development and manufacturing. Collect icIEF peaks directly rather than resorting to the resource-intensive ion exchange chromatography (IEX) for fractionation.
Regulatory Filing
Investigate icIEF fractions by peptide mapping and potency assays. Have more confidence in the modifications underlying each icIEF peak. Accelerate timelines by avoiding weeks/months of IEX method development and bridging studies. Bring forward novel biotherapeutics that were previously too challenging to characterize by IEX.
Manufacturing
Rapid and thorough peptide mapping of new and unexpected icIEF peaks. Simplify the scale-up process. Understand inconsistencies at different manufacturing sites. Quickly diagnose issues that arise during production. Minimize the risk of product quality concerns and manufacturing delays.
Process Development
Run peptide mapping on icIEF peaks under a variety of changing conditions. Optimize drug performance in terms of formulation, stability, process development, and more. Develop a thorough understanding of the drug compound. Avoid surprises and unnecessary delays during regulatory filing and manufacturing.
Milligram-Scale icIEF Fractionation: Sharpening In-Depth Characterization of Protein Charge Heterogeneity
The CEInfinite icIEF fractionation system from Advanced Electrophoresis Solutions Ltd. (AES) significantly accelerates comprehensive insights into charge variant modifications during biopharma development, helping to mitigate costly setbacks related to scale-up and regulatory compliance.
CEInfinite preparative icIEF is advancing the evolution of icIEF platforms by integrating cutting-edge instrument design, high-resolution critical reagents, and a standardized workflow. Its exceptional resolution in icIEF separation, paired with unmatched run-to-run repeatability for icIEF fractionation, guarantees superior performance with high throughput, purity, and yield. This enables effective solutions to stability, manufacturing, safety, and efficacy concerns.
Utilizing custom-designed capillary cartridges offering ultra-high preparation capacity, the system effortlessly delivers milligram-scale yields within a week, facilitating detailed post-fractionation characterization. By integrating with LC-MS, IEX-MS, and MALDI-TOF, the CEInfinite system supports analysis at both the intact mass and peptide mapping levels. Additionally, its combination with SPR and potency assays further extends the application of icIEF fractionation into bioactivity studies.
icIEF Method Transfer
(1-3 days)
Directly transfer your icIEF method or improve resolution with narrow-range AESlytes® and CEInfinite coatings.
Method Development
(1-5 days)
Define each fraction interval size, the initial sample concentration, and the mobilization speed.
Auto Fractionation
(~50-250 µg/day)
20-30 runs/day at 4-24 µg of sample per run followed by buffer exchange to remove ampholytes and more.
Instrument Spare Parts
Technical Notes & Publications
CEInfinite Auto icIEF Fractionation Workflow
Fractionate Faster. Analyze Deeper.
Eliminate bottlenecks in icIEF fractionation with automated, high-yield peak collection. Let’s talk about how CEInfinite can accelerate your workflow.? Get in touch!
