Product Description
Hydrophobic Interaction Chromatography (HIC) HPLC is well suited for the analysis of proteins and antibody–protein bioconjugates based on their hydrophobicity. While the underlying principle is similar to reverse-phase chromatography, HIC employs a lower degree of hydrophobic substitution on the stationary phase, allowing compounds to be eluted under more aqueous, less denaturing conditions.
HIC is widely used for analyzing drug loading and heterogeneity in antibody–drug conjugates (ADCs). For ADCs prepared via reduced thiol conjugation, HIC HPLC can also be used to estimate the drug-to-antibody ratio (DAR). Antibodies with the same number of conjugated drug molecules (i.e., identical DAR) exhibit similar hydrophobicity and typically elute as a single peak. In contrast, ADC samples such as MMAE conjugates often display multiple peaks corresponding to species with varying drug loads.
Following analysis at CellMosaic, a standard PDF report of the HPLC profiles will be provided.
Pricing is per sample. A $250 setup fee applies per batch analysis and will be waived if the conjugate is prepared using a CellMosaic kit or if an extended turnaround time (up to 9 business days) is acceptable. Standard turnaround time is typically 1–3 business days.
Sample submission for biopolymer, bioconjugate, and ADC:
1) For biopolymers, bioconjugates, and ADCs, dilute samples in PBS buffer to 0.5–2 mg/mL. Centrifuge (≥5000 × g) to remove particulates. Transfer a minimum of 50 μL of the supernatant to a 500 μL screw-cap microcentrifuge tube.
2) Please download and complete the Sample Submission Form with all required information. Email a copy, and include a printed copy with your samples. Ship with a cold pack via overnight or earliest delivery.
For analysis of bioconjugates such as antibody drug conjugates, always ship a sample of un-labeled biopolymer for analysis at the same time.
Related Products:
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.
