Trace Detection of Antibiotic Impurities in Cell Culture

Impurities derived from cell culture include but are not limited to inducers, antibiotics, serum and other media components. These impurities need to be tested and evaluated one by one using risk assessment and risk management methods. If there is a potential impact on product safety, the removal of such impurities to an acceptable low level in the downstream purification process may require verification, or final product testing and establishment of specification limits.

Process-related impurities come from the manufacturing process itself and can be divided into three categories: cell matrix derivation, cell culture derivation and downstream derivation. On the other hand, contaminants are unwanted materials, such as foreign viruses, which are introduced into the manufacturing process inadvertently.

a) Impurities derived from cell substrates include, but are not limited to, proteins and nucleic acids derived from host organisms (host cell genome, vector or total DNA). For host cell proteins, sensitive assays (such as immunoassays) capable of detecting various protein impurities are usually used. In the case of immunoassays, the polyclonal antibodies used for testing are usually collected by immunizing animals with an appropriate preparation from production cells (eg, filtered harvest, partially purified). The level of DNA from host cells can be detected by direct analysis of the product (such as qPCR, immunoenzyme technology). Clearance studies may include small-scale spike experiments to demonstrate the removal of cell matrix-derived impurities, such as nucleic acids and host cell proteins, and can sometimes be used to eliminate the need to establish acceptance criteria for these impurities.

b) Impurities derived from cell culture include but are not limited to inducer antibiotics, serum and other medium components. These impurities need to be tested and evaluated on a case-by-case basis using risk assessment and risk management methods. In the case of potential impact on product safety, it may be necessary to verify the removal of such impurities to an acceptable low level in the downstream purification process, or to establish final product testing and specification limits.

c) Impurities from downstream sources include but are not limited to enzymes, chemical and biochemical processing reagents (such as guanidine, dyes, oxidants and reducing agents), inorganic salts (such as heavy metals, non-metal ions), solvents, carriers, ligands (such as protein A) ) And other leachables. For impurities derived from cell culture, risk assessment and risk management methods should be used to assess these impurities on a case-by-case basis. Where appropriate, consider developing analytical methods for these impurities and verify their removal.

Effective removal of process-related impurities or process residues is very important for pharmaceutical and biopharmaceutical development. Manufacturers need to have a good understanding of process residues in the early stages of development in order to better optimize the production process and establish acceptance criteria for ongoing testing requirements (such as batch release testing). These process-related impurities are usually present in challenging sample matrices in trace amounts, so quantification can be challenging.

Among cell culture-derived impurities, antibiotics are common cell culture-derived impurities in the biological production process. Antibiotic testing is necessary to ensure that the purification process effectively removes these chemicals, keeps them at or below the maximum allowable levels, and complies with any product-specific regulatory guidelines. Creative Diagnostics has extensive experience in the development of methods for the detection and validation of trace antibiotics (such as kanamycin, tobramycin, gentamicin, and chloramphenicol). In order to meet the requirements for monitoring and controlling process-related impurities and contaminants in the biological production process described in the ICH Q6B Biomolecule Production Guidelines, we have developed a set of highly sensitive antibiotic ELISA kits to analyze antibiotic residues. These kits are suitable as good impurity detection tools used in the biological production process.