0010057AE) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). A1148301), and anti-clumping agent (catalog no. The data was obtained from an off-line UV-Vis spectrum of samples from lab-scale CHO cell culture supernatants.ĭynamis (catalog no. In this study, the potential of monitoring VCD and antibody titers from lab-scale Chinese hamster ovary (CHO) cell cultures by applying UV-Vis spectra in combination with MVDA and PLS calibration model was demonstrated. There has been a previous report on successful application of this technique to monitor glutamine and glucose concentrations in BHK-21 cell cultures. However, this method is more attractive in terms of ease of handling and cost, which is more suitable for lab-scale or manufacturing of therapeutic antibody in low-to-middle-income countries. Ultraviolet-Visible (UV-Vis) spectra provide less information when compared with other spectroscopy techniques as the bands are less specific and have high chances of spectral overlapping. This is because these spectroscopic methods can be used to identify chemical functional groups. The most popular spectroscopies combined with MVDA in PAT tool are Raman spectroscopy, near-infrared spectroscopy (NIR), and Fourier-transform infrared spectroscopy (FTIR). Recently, multivariate data analysis (MVDA) such as principal component analysis (PCA) and partial least squares (PLS), in conjunction with spectroscopic spectra, has been considered to be a powerful tool in the process analytical technology (PAT) or quality by design (QbD) of biopharmaceutical processes. For example, the determination of antibody titers by ELISA involves several reagents and a microplate reader, and the determination of %viability by trypan blue staining requires a cell counter or inverted microscope as well as experienced personnel. However, monitoring of these parameters need specific methods and instruments, which are expensive, laborious, require substantial volume of samples, or sophisticated instruments in case of real-time measurement. Among these parameters, VCD and antibody titers are the two key parameters for determining optimal condition for cell line cultivation during bioprocess development. These bioprocess parameters are necessary to monitor the performance of cell lines both during the process development and routine manufacturing. Multiple bioprocess parameters such as viable cell density (VCD), antibody titer, pH, temperature, %CO 2, and level of metabolites need to be tightly monitored in order to obtain maximum yield and to ensure consistency of product quality. Since the production of therapeutic antibodies involves the use of living cells, the manufacturing process is much more complicated than generating small molecule drugs, which requires chemical reactions. These results indicated that UV-Vis combined with PLS has potential to be used for monitoring antibody titer, VCD, and % cell viability for both online and off-line therapeutic production process. The results indicated that while all predicted % cell viability were very similar to the actual value at RSEP value of 6.7 and R 2 of 0.8908, the predicted productivity from 14 of 18 samples were closed to the reference measurements at RSEP value of 22.4 and R 2 of 0.8522. Then, the percentage of cell viability and productivity were predicted from a set of samples of polyclones. The results indicated that the optimal PLS models could be used to predict antibody titer and VCD with the linear relationship between reference values and predicted values at R 2 values ranging from 0.8 to > 0.9 in supernatant samples obtained from four different single clones and in polyclones that were cultured in various selection stringencies. In this study, we demonstrated that as low as 2 μL of culture supernatants were sufficient for the analysis using UV-Vis spectrum assisted with partial least squares (PLS) model. Typically, determination of each parameter requires 10–100 μL of supernatant sample, which is not suitable for small scale cultivation. Antibody titer and viable cell density (VCD) are two important parameters that need to be closely monitored during the process of cell line development and manufacturing of therapeutic antibodies.
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