PG13 Host Cell Protein ELISA Kit

Sample

Expression products

Detection Method

sELISA

Intended Use

This kit is intended for use in determining the presence of host cell protein impurities in products manufactured by expression in PG13 cells. The kit is for Research and Manufacturing Use Only and is not intended for diagnostic use in humans or animals.

Contents of Kit

1. Anti-PG13: HRP
Affinity purified goat antibody conjugated to HRP in a protein matrix with preservative. 1×12mL
2. Anti-PG13 coated microtiter strips
12×8 well strips in a bag with desiccant
3. PG13 HCP Standards
PG13 HCPs in bovine serum albumin with preservative. Standards at 0, 3, 6, 12, 25, 50, 100 and 200ng/mL. 1 mL/vial
4. Stop Solution
0.5M sulfuric acid. 1×12mL
5. TMB Substrate
3,3',5,5' Tetramethylbenzidine. 1×12mL
6. Wash Concentrate (20×)
Tris buffered saline with preservative. 1×50mL

Storage

1. All reagents should be stored at 2°C to 8°C for stability until the expiration date printed on the kit.
2. After prolonged storage, you may notice a salt precipitate and/or yellowing of the wash concentrate. These changes will not impact assay performance. To dissolve the precipitate, mix the wash concentrate thoroughly and dilute as directed in the 'Reagent Preparation' section. Reconstituted wash solution is stable until the expiration date of the kit.

General Description

Retroviral packaging in PG13 cells is a cost-effective way to produce a high titer of retrovirus capable of infecting human cells for gene and cell therapies. The manufacturing and purification process of these products leaves the potential for impurities by host cell proteins (HCPs) from PG13 cells. Such impurities can reduce the efficacy of the therapeutic agent and result in adverse toxic or immunological reactions and thus it is desirable to reduce HCP impurities to the lowest levels practical. This simple to use, objective, and semi-quantitative ELISA is a sensitive and specific method to aid in optimal purification process development, process control, and in routine quality control and product release testing. This kit is "generic" in the sense that it is intended to react with essentially all of the HCPs that could contaminate the product independent of the purification process. The antibodies have been generated against and affinity purified using a mild lysate of PG13 cells. The antibodies used in this kit were characterized by Antibody Affinity Extraction (AAE) and Mass Spectrometry, demonstrating reactivity to the majority of HCPs. Special procedures were utilized in the generation of these antibodies to ensure that low molecular weight and less immunogenic impurities as well as high molecular weight components would be represented. As such, this kit can be used as a process development tool to monitor the optimal removal of host cell impurities as well as in routine final product release. This highly sensitive ELISA kit was qualified for testing of final product HCPs by using actual in-process and final drug substance samples. Each user of this kit is encouraged to perform a similar qualification study to demonstrate it meets their analytical needs. The suitability of this kit for a given sample type and product must be determined and qualified experimentally by each laboratory.