Reversed-Phase Analyses of Monoclonal Antibodies Using High Temperatures
The Application Notebook
This application note shows how to achieve robust chromatographic results for two commercially available mAbs: Adalimumab (Humira®) and Bevacizumab (Avastin®).
Monoclonal antibodies (mAbs) are immunological active proteins, which bind specifically to certain cells or proteins. This will stimulate the immune system to attack those targets.
mAbs are very important for the treatment of different kinds of cancer and autoimmune diseases. Nowadays, a broad variety of therapeutical antibodies are available on the market and several more are in research and development.
Due to their molecular weight of about 150 kDa, intact antibodies are usually analyzed by IEX, SEC, or HIC. In addition, reversedâphase methods are an easy tool as well.
However, a lack of sensitivity and resolution has been a hurdle in the past. With modern reversed phases addressing the requirements of these analytes, it is easy to find a suitable method.
Successful analysis in reversed-phase mode for mAbs is enhanced by employing a temperature stable (> 60 °C), widepore stationary phase.
This application note shows how to achieve robust chromatographic results for two commercially available mAbs: Adalimumab (Humira®) and Bevacizumab (Avastin®).
The increase in temperatures leads to higher sensitivity and a sharper peak of Adalimumab at temperatures >60 °C (Figure 1).
Bevacizumab shows robust results starting from 70 °C, whereas at lower temperatures no peak was detected (Figure 2).
With YMC-Triart Bio C4, elevated temperatures can easily be applied, due to stability up to 90 °C. In addition, the surface comprising 30 nm/300 Å is beneficial for resolution. In combination with a wide pH range of 1–10 YMC-Triart Bio C4 is a well-suited tool for any mAb (U)HPLC method.
YMC Europe GmbH
Tel.: +49 2064 4270
E-mail: info@ymc.de
Website: www.ymc.de
AI and GenAI Applications to Help Optimize Purification and Yield of Antibodies From Plasma
October 31st 2024Deriving antibodies from plasma products involves several steps, typically starting from the collection of plasma and ending with the purification of the desired antibodies. These are: plasma collection; plasma pooling; fractionation; antibody purification; concentration and formulation; quality control; and packaging and storage. This process results in a purified antibody product that can be used for therapeutic purposes, diagnostic tests, or research. Each step is critical to ensure the safety, efficacy, and quality of the final product. Applications of AI/GenAI in many of these steps can significantly help in the optimization of purification and yield of the desired antibodies. Some specific use-cases are: selecting and optimizing plasma units for optimized plasma pooling; GenAI solution for enterprise search on internal knowledge portal; analysing and optimizing production batch profitability, inventory, yields; monitoring production batch key performance indicators for outlier identification; monitoring production equipment to predict maintenance events; and reducing quality control laboratory testing turnaround time.
