title: Clearance of extractables and leachables from single-use technologies via ultrafiltration/diafiltration operations.
abstract: Quantifying the clearance of extractables and leachables (E/L) throughout ultrafiltration/diafiltration (UFDF) operations allows for greater flexibility in the implementation of single-use technologies in steps upstream of the UFDF process. A proof-of-concept study was completed in which the clearance of 7 E/L from single-use technologies (trimethylsilanol, hexanoic acid, butyrolactone, t-butyl alcohol, caprolactam, acetonitrile, and benzyl alcohol) in four representative proteins were measured and monitored during the UFDF process using quantitative NMR. This study demonstrated that the defined E/L spiked into a variety of protein solutions can be cleared to <1 ppm by 9 diavolumes from a maximum initial load concentration of 1,000 ppm. However, in some cases a rebound effect was observed in the recovered pool to >1 ppm, which is explained in detail. The overall clearance trend observed for both buffer control and protein-containing solutions resembled the ideal clearance trend where no apparent interactions were observed between E/L with the protein, UFDF system, or with other defined E/L which may be present in the system. Additionally, the UFDF system is capable of clearing these potential E/L from single-use technologies below 1 ppm irrespective of initial concentrations in the load (1,000 or 100 ppm), independently from the type of protein. In general, mass recoveries were within ±15% of each spiked compound in protein solutions and their respective buffer controls, suggesting spiked E/L do not interact strongly with protein. By demonstrating the product independent clearance trends of the spiked E/L across UFDF, these results will contribute to the simplification of the E/L toxicology assessment and allow modular manufacturing approach for single-use technologies in biopharmaceutical manufacturing. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:718-724, 2016.
authors.affiliation: Genentech, Inc, South San Francisco, CA, 94080.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Magarian
authors.affiliation: Genentech, Inc, South San Francisco, CA, 94080.
authors.forename: Kate
authors.initials: K
authors.lastname: Lee
authors.affiliation: Genentech, Inc, South San Francisco, CA, 94080.
authors.forename: Kunal
authors.initials: K
authors.lastname: Nagpal
authors.affiliation: Genentech, Inc, South San Francisco, CA, 94080.
authors.forename: Ken
authors.initials: K
authors.lastname: Skidmore
authors.affiliation: Genentech, Inc, South San Francisco, CA, 94080.
authors.forename: Ekta
authors.initials: E
authors.lastname: Mahajan
mesh_terms: D011506:Proteins / Q000032:analysis*; D014462:Ultrafiltration
chemical_list: D011506:Proteins
keywords: NMR; clearance; extractables and leachables; single-use; ultrafiltration/diafiltration