Reverse Phase Liquid Chromatography (RPLC)

Reversed phase liquid chromatography (RPLC) is a method that separates components based on their hydrophilic or hydrophobic nature. Benefiting from its high resolution and ability to be coupled with other techniques, this technique has found wide application in antibody drug characterization. These advanced technologies and methods can overcome the challenges of antibody drug characterization and generate enhanced information for better characterization and understanding of antibody drug therapy products. Creative Proteomics provides high-resolution reversed-phase liquid chromatography technology and comprehensive analytical services to antibody drug researchers and developers worldwide. Our broad portfolio of technologies and services provides valuable insights and solutions to ensure the highest quality and efficiency in the antibody drug development process.

Application of RPLC in Characterization of Antibody Drugs

Reversed-phase liquid chromatography (RPLC) is one of the most versatile/popular chromatographic techniques for pharmaceutical analysis, providing excellent sensitivity, selectivity, accuracy, and robustness. The technique is often combined with mass spectrometry (MS) to provide more accurate and high-quality characterization data on the chemical and structural integrity of antibody drugs. Its wide range of characterization capabilities include:

• Primary structural characterization of antibody drugs.
• Isolation and characterization of impurities/degradation products.
• Commonly used in peptide mapping experiments to assess post-translational modifications (PTM) of antibody drugs, including oxidation, deamidation, truncation/clipping, isomerization, disulphide bond integrity, etc.
• For ADC analysis, when interest lies in the ADC as a whole, RPLC can be used to determine isomeric distribution, compare conjugate heterogeneity, and conduct peptide mapping. Whereas when the focus is on the small molecule side, RPLC can be utilized for free drug analysis, residual solvent analysis, as well as to study the stability of the small molecule drugs.

Advantages of Reversed-Phase Liquid Chromatography

Reversed-phase liquid chromatography (RPLC) offers several advantages for antibody drug characterization, making it a widely used technique in this field. These advantages include:

• Versatility: RPLC can handle a wide variety of samples and can separate and analyze various types of antibody therapeutic molecules.
• Flexibility: The selection of stationary phase, mobile phase and detection technique is flexible according to specific assay requirements.
• Stability: The high stability of RPLC ensures separation efficiency and consistent, reliable characterization results.
• High separation efficiency: The narrow peaks obtained in RPLC ensure accurate quantification and characterization of antibody drug samples.
• MS compatibility: RPLC, when coupled with mass spectrometry (MS), provides detailed information on the molecular weight, post-translational modifications, fragmentation patterns, and other structures of the antibody.

Our Capabilities

• Advanced RPLC techniques

A variety of different RPLC methods used for antibody drug characterization such as gradient elution, isocratic elution and in combination with other techniques to provide comprehensive characterization data.

• Method development and validation

Our experienced characterization experts develop robust RCPL separation methods based on customer needs, including customizing optimal mobile and stationary phases for efficient separation and resolution, optimizing gradient elution, sample loading and temperature conditions. Our system supports method development on a wide range of stationary phases, ensuring flexibility in achieving desired selectivity and resolution for different antibody drugs. All of our assays are fully validated to ensure reliability and accuracy.

• Comprehensive data analysis

We provide sophisticated data analysis software that enables efficient integration, peak identification, and quantitation, facilitating comprehensive characterization of antibody drugs.

Creative Proteomics is committed to providing advanced technology and expertise to our partners in antibody drug development. Our RPLC systems ensure robust and reproducible results, minimizing errors and increasing confidence in the data obtained. This is critical for key decisions in antibody drug development and manufacturing. Contact us to learn more about our service and we will be happy to serve you.