After synthesizing the ADC, I ran HIC and observed a peak. Does this indicate successful coupling?
After the synthesis of antibody-drug conjugates (ADC), a new peak was observed using hydrophilic interaction chromatography (HIC), which indeed may indicate that the conjugation reaction has occurred. HIC can help distinguish substances with different hydrophilic or hydrophobic characteristics. If the drug in the ADC successfully conjugates with the antibody, its hydrophilicity or hydrophobicity will change, typically resulting in a new chromatographic peak in HIC analysis.
However, the appearance of a new peak alone cannot confirm that the conjugation was entirely successful. To verify whether the ADC conjugation was successful and to determine the degree and consistency of the conjugation, you may need to perform additional analyses involving the following steps:
1. Mass spectrometry (MS):
Used to confirm the molecular weight of the new peak and compare it with the theoretical drug-antibody conjugate to confirm the presence of the conjugate.
2. Conjugation efficiency detection:
Evaluate the conjugation rate of the drug to the antibody in the ADC using quantitative methods such as ELISA or fluorescence labeling.
3. Bioactivity testing:
Ensure that the conjugated ADC retains the expected bioactivity.
Bio-tech Pack Biological Technology -- A quality service provider in biopharmaceutical characterization and multi-omics mass spectrometry detection.
Related services:
Antibody-drug conjugates (ADCs) Analysis
Protein structure identification
Post-translational modification proteomics analysis
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