Evaluation of hydrophobic‐interaction chromatography resins for purification of antibody‐drug conjugates using a mimetic model with adjustable hydrophobicity
| Autor: | Manuela Sevilla, Egbert Müller, Patrick Endres |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Drug
Immunoconjugates Ph dependent solubility Resolution (mass spectrometry) purification media_common.quotation_subject Filtration and Separation 030226 pharmacology & pharmacy Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine fluorescein 5‐isocyanate Humans Fluorescein Cytotoxic molecules Chromatography High Pressure Liquid 030304 developmental biology media_common Liquid Chromatography 0303 health sciences drug antibody ratio Chromatography biology antibody drug conjugates Chemistry Hydrophilic interaction chromatography hydrophobic interaction chromatography Resins Synthetic Models Chemical biology.protein Antibody Hydrophobic and Hydrophilic Interactions Conjugate Research Article |
| Zdroj: | Journal of Separation Science |
| ISSN: | 1615-9314 1615-9306 |
| Popis: | Antibody drug conjugates are cytotoxic pharmaceuticals, designed to destroy malignant cells. A cytotoxic molecule is attached to an antibody that binds specific to a cancer‐cell surface. Given the high toxicity of the drugs, strict safety standards have to be kept. For this reason, an antibody drug conjugates model was developed with fluorescein 5‐isothiocyanate as the nontoxic payload surrogate. Due to the similar hydrophobicity, this model is used to establish a suitable purification process and characterization method for antibody drug conjugates. Because of the pH dependent solubility of fluorescein, the hydrophobicity of conjugates can be modulated by the pH value. Based on the complex heterogeneity and hydrophobicity of the conjugates a chromatographic purification is challenging. Hydrophobic interaction chromatography is used for analytical as well as for preparative separation. Because of the increased hydrophobicity of the conjugates compared to native antibody, hydrophobic interaction chromatography often suffer from resolution and recovery problems. Conjugates were separated differing on the number of payloads attached to the antibody. For this matter, the drug–antibody ratio is determined and used as a quantitative term. The conjugates are purified at high recoveries and resolution by step gradients using suitable resins, allowing the separation of the target drug–antibody ratio. |
| Databáze: | OpenAIRE |
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