Quantitative evaluation of colloidal stability of antibody solutions using PEG-induced liquid-liquid phase separation.
نویسندگان
چکیده
Colloidal stability of antibody solutions, i.e., the propensity of the folded protein to precipitate, is an important consideration in formulation development of therapeutic monoclonal antibodies. In a protein solution, different pathways including crystallization, colloidal aggregation, and liquid-liquid phase separation (LLPS) can lead to the formation of precipitates. The kinetics of crystallization and aggregation are often slow and vary from protein to protein. Due to the diverse mechanisms of these protein condensation processes, it is a challenge to develop a standardized test for an early evaluation of the colloidal stability of antibody solutions. LLPS would normally occur in antibody solutions at sufficiently low temperature, provided that it is not preempted by freezing of the solution. Poly(ethylene glycol) (PEG) can be used to induce LLPS at temperatures above the freezing point. Here, we propose a colloidal stability test based on inducing LLPS in antibody solutions and measuring the antibody concentration of the dilute phase. We demonstrate experimentally that such a PEG-induced LLPS test can be used to compare colloidal stability of different antibodies in different solution conditions and can be readily applied to high-throughput screening. We have derived an equation for the effects of PEG concentration and molecular weight on the results of the LLPS test. Finally, this equation defines a binding energy in the condensed phase, which can be determined in the PEG-induced LLPS test. This binding energy is a measure of attractive interactions between antibody molecules and can be used for quantitative characterization of the colloidal stability of antibody solutions.
منابع مشابه
Evaluation of effects of pH and ionic strength on colloidal stability of IgG solutions by PEG-induced liquid-liquid phase separation.
Colloidal stability of IgG antibody solutions is important for pharmaceutical and medicinal applications. Solution pH and ionic strength are two key factors that affect the colloidal stability of protein solutions. In this work, we use a method based on the PEG-induced liquid-liquid phase separation to examine the effects of pH and ionic strength on the colloidal stability of IgG solutions. We ...
متن کاملSeparation of Somatropin and Its Degradation Products by High-Performance Liquid Chromatography Using a Reversed-Phase Polymeric Column
The accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. In this study, a gradient RP-HPLC method is described for the separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min flow rate of a linear gradient of 0.1% v/...
متن کاملExtraction of Silver Ions from Aqueous Solutions by Emulsion Liquid Membrane
A comprehensive study pertaining to the emulsion liquid membrane (ELM) extraction process to enrich dilute aqueous solutions of silver salt is presented. The study has highlighted the importance and influence of membrane composition for maximizing the extraction of Ag+ ions. The liquid membrane was made up of Cyanex-302 as an extractant and the industrial solvent mainly consists of p...
متن کاملNano composite PEBAX®/PEG membranes: Effect of MWNT filler on CO2/CH4 separation
The performances of two-phase polymer-liquid PEBAX®/polyethylene glycol (PEG) and three-phase polymer-liquid-solid PEBAX®/PEG/MWNT thin film composite membranes for CO2 and CH4 permeation were studied. The effect of temperature and MWNT/PEBAX® ratio on single gas (CO2 and CH4) permeability was investigated. The permeat...
متن کاملSeparation of Somatropin and Its Degradation Products by High-Performance Liquid Chromatography Using a Reversed-Phase Polymeric Column
The accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. In this study, a gradient RP-HPLC method is described for the separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min flow rate of a linear gradient of 0.1% v/...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Molecular pharmaceutics
دوره 11 5 شماره
صفحات -
تاریخ انتشار 2014