Isoelectric Focusing Electrophoresis (iCIEF) Service

In the development and quality control of protein-based therapeutics, understanding charge heterogeneity is essential. Creative Biostructure offers advanced isoelectric focusing electrophoresis (iCIEF) services, providing precise analysis of protein charge variants to support drug development, biosimilar characterization. Our expertise in isoelectric focusing electrophoresis ensures high-resolution separation, accurate pI determination, and comprehensive data interpretation for biopharmaceutical applications.

Why is Isoelectric Focusing Electrophoresis Essential?

Isoelectric focusing electrophoresis (IEF) is a precise analytical technique used to separate proteins based on their isoelectric points (pI)—the pH at which a protein has no net charge. This property directly influences protein structure, stability, and function, making IEF crucial for biopharmaceutical research and drug quality control.

iCIEF, an advanced form of IEF, offers higher resolution, faster method development, superior sensitivity, and improved reproducibility compared to traditional capillary isoelectric focusing (CIEF). By applying an electric field within a capillary column, iCIEF establishes a stable pH gradient that enables proteins to migrate and focus at their respective pI values, providing high-resolution data for charge heterogeneity analysis.

Our iCIEF Services at Creative Biostructure

As a leader in structural biology research, Creative Biostructure offers comprehensive iCIEF services tailored to the needs of the pharmaceutical and biotechnology industries. Our services play a critical role in protein drug development, biosimilar characterization, and quality control.

Service Workflow of iCIEF Service

1. Capillary Filling

The first step in iCIEF involves carefully filling the capillary with a precisely selected ampholyte mixture. This solution establishes the foundation for a stable pH gradient, ensuring optimal conditions for protein separation.

2. Establishing pH Gradient

A stable and reproducible pH gradient is created within the capillary to allow proteins to migrate to their isoelectric points. The selection of appropriate ampholytes and control of gradient stability are crucial for achieving high-resolution separation.

3. Sample Loading

Protein samples are introduced into the capillary under controlled conditions to prevent degradation or aggregation. Proper sample preparation and loading techniques ensure reliable and reproducible results.

4. Protein Migration

Once an electric field is applied, proteins migrate through the capillary and focus at their respective isoelectric points. This process results in distinct bands corresponding to different charge variants, allowing precise characterization of protein heterogeneity.

5. Protein Detection

Real-time imaging and detection technologies capture high-resolution electropherograms, enabling accurate visualization of charge variant distribution. Sensitive detection methods enhance the reliability of the analysis.

6. Data Analysis

Advanced computational tools are used to interpret electrophoresis data, including pI determination and quantitative analysis of charge variants. Our experts provide comprehensive reports with detailed insights into protein charge heterogeneity.

Figure 1. iCIEF Service Process. (Creative Biostructure)

Why Choose Creative Biostructure?

• High-Precision Charge Variant Analysis: Utilizing advanced iCIEF technology, we ensure accurate characterization of protein charge heterogeneity.
• Rapid Method Development: Our experts quickly customize optimized analytical methods, accelerating your research timelines.
• Comprehensive Technical Support: Our team provides expert consulting and data interpretation to ensure meaningful insights from your results.
• Stringent Quality Control: We adhere to rigorous analytical standards to guarantee consistent and reliable data for regulatory submissions.
• Flexible & Customizable Solutions: Tailored services based on specific project requirements, accommodating a wide range of biopharmaceutical applications.

Frequently Asked Questions

• What types of samples are suitable for iCIEF analysis?

  Our iCIEF service supports the analysis of peptides, proteins, nucleic acids, and other biomolecules requiring precise charge variant characterization.

• What information does iCIEF analysis provide?

  iCIEF offers detailed insights into the charge variant distribution of a protein, including qualitative and quantitative data on different isomeric forms.

• How does iCIEF compare to traditional CIEF?

  iCIEF offers higher resolution, increased sensitivity, faster analysis times, and improved reproducibility compared to conventional capillary isoelectric focusing techniques.

• Why is charge variant analysis important for protein therapeutics?

  Charge variants can impact protein stability, efficacy, and immunogenicity. Understanding these variations is essential for ensuring batch-to-batch consistency and regulatory compliance.

• Can iCIEF be used for biosimilar comparison studies?

Yes, iCIEF is a critical tool for comparing biosimilars to reference biologics, helping to establish structural and functional equivalence.

For customized iCIEF solutions, contact us today to discuss your project requirements and obtain a detailed quote.

Ordering Process

References

1. Mack S, Arnold D, Bogdan G, et al. A novel microchip‐based imaged CIEF‐MS system for comprehensive characterization and identification of biopharmaceutical charge variants. Electrophoresis. 2019, 40(23-24): 3084-3091.
2. Zhang X, Kwok T, Zhou M, et al. Imaged capillary isoelectric focusing (icIEF) tandem high resolution mass spectrometry for charged heterogeneity of protein drugs in biopharmaceutical discovery. Journal of Pharmaceutical and Biomedical Analysis. 2023, 224: 115178.