Hydrophobic Interaction Chromatography (HIC) HPLC is good for analyzing proteins or antibody/protein bioconjugates based on their hydrophobicity. The theory behind HIC is similar to reverse-phase chromatography; however, the degree of substitution of HIC adsorbents is diluted compared to reverse-phase chromatography. Thus, the compound can be eluted in a more polar and less denaturing environment.
HIC has been used extensively to analyze drug loading in antibody drug conjugates (ADC) and their heterogeneity. For ADCs prepared via a reduced thiol of the antibody, HIC HPLC can be further used to calculate the DAR. Antibodies loaded with the same number of drugs (same DAR) will have similar hydrophobicity and be eluted as a single peak. For a typical MMAE ADC, multiple peaks represent various amounts of drug-loaded ADCs.
After submitting samples to CellMosaic for analysis, a standard pdf file of the HPLC profiles will be supplied.
Per sample charge.
Setup fee of $250 applies per batch analysis. Setup fee will be waived if the conjugate is made using CellMosaic's kit or if sample can be analyzed within a longer timeframe of up to 7 business days. Standard services is 1-3 business days.
Sample submission for biopolymer, bioconjugate, and ADC:
1) Dilute your samples in PBS buffer to 1 mg/mL, and then transfer minimum 50 μL of the diluted solution to a 500 μL micro-centrifuge tube. Label the vial properly.
2) Ship your samples with a cold pack for overnight delivery.
For analysis of bioconjugates such as antibody drug conjugates, always ship a sample of un-labeled biopolymer for analysis at the same time.
HIC Buffer Set (CM02025): CellMosaic® offers this HIC buffer set for our customers to use with any HIC column. The CM02025 product sheet contains all of the information and methodology needed to run an HIC HPLC analysis.