title: Quantitation of residual trypsin in cell-based therapeutics using immobilized α-1-antitrypsin or SBTI in an ELISA format.
abstract: An Enzyme-Linked Immunosorbent Assay (ELISA) has been developed for the quantitation of porcine trypsin as a process residual in cell therapy products based on its capture by either of two immobilized anti-trypsins, α-1-antitrypsin (α1AT) or soybean trypsin inhibitor (SBTI) followed by detection with a polyclonal goat anti-porcine trypsin-IgG conjugated with peroxidase. It was demonstrated that an extended range of antigen quantitation could be achieved that covered nearly three orders of magnitude of trypsin concentration. The utility of the assay was demonstrated by its application to samples generated in a cell-based therapeutic manufacturing setting.
authors.affiliation: BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States.
authors.forename: James A
authors.initials: JA
authors.lastname: Braatz
authors.affiliation: BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States.
authors.forename: Christopher
authors.initials: C
authors.lastname: Elias
authors.affiliation: BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States.
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Finny
authors.affiliation: BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States.
authors.forename: Huan
authors.initials: H
authors.lastname: Tran
authors.affiliation: BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States. Electronic address: michael.mccaman@lonza.com.
authors.forename: Michael
authors.initials: M
authors.lastname: McCaman
mesh_terms: D000818:Animals; D064987:Cell- and Tissue-Based Therapy; D004797:Enzyme-Linked Immunosorbent Assay; D055767:Immobilized Proteins / Q000737:chemistry; D013234:Stem Cells / Q000737:chemistry*; D013552:Swine; D014357:Trypsin / Q000032:analysis*; D014361:Trypsin Inhibitors / Q000737:chemistry*; D000515:alpha 1-Antitrypsin / Q000737:chemistry*
chemical_list: D055767:Immobilized Proteins; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; D014357:Trypsin
keywords: Cell-based therapeutic; ELISA; Porcine trypsin; Process residual; α-1-Antitrypsin