Establishment of a rapid methodology for quantitative monitoring of monoclonal antibodies production

Introduction: Monoclonal antibodies (mAb) are widely used in areas such as research, diagnostics, and therapeutic solutions for cancer and immune disorders.Quantification during mAb production is challenging due to the numerous components and conditions used in the process.Process analytical monitoring is therefore important to understand the state of the process allowing the maintenance of the desired state.Thereby, it is important to quantify mAb concentration quickly and accurately during the production process, in different media, from crude biological material to drug product.Objectives: To establish a rapid and accurate methodology to determine mAb content in processes that can be implemented in process analytical technology system. Methodology:The mAb concentration was determined by high-performance liquid chromatography on protein A affinity analytical column with spectrophotometric detection in the UV range.For this purpose, an analytical curve was constructed determining its linear range, detection (DL) and quantification (QL) limit, precision, repeatability, intermediate precision, accuracy and selectivity.To evaluate matrix interference, a standard addition curve was done using crude material to mimic a complex condition. Results:The study of linearity in the range of 0.050 to 0.250 mg mL-1 showed a coefficient of determination of 0.999.No matrix effect was evidenced, with a relative standard deviation of 0.5% between the angular coefficients of the curves.The recovery was between 98.0 and 100.9% with no statistical difference of 1 (100%).The obtained DL and QL were 0,0036 and 0,011 mg mL-1, calculated from the analytical curve. Conclusion:The present method proved applicable for analysis in process and has the capability to be used in different stages of the production process.This development is possibility well suited as a platform method using affinity chromatography in a gradient of less than 5 min, ensuring direct application in Quality and Process Control routines.By using this method, it will be possible to increase sample processing capacity in a fast, accurate and reproductive way.