Description

The expression of viral vectors and vaccines in Vero cells is a cost-effective method for the production of commercial quantities of a drug. The manufacturing and purification process of these products leaves the possibility of impurities by the HCPs in the Vero cells. Such impurities may reduce the efficacy of the therapeutic or vaccine agent and result in adverse toxic or immunological reactions. Therefore, it is desirable to reduce HCP impurities to the lowest practical levels.

This kit was developed using antibodies that have been generated and affinity purified using mild Vero cell lysate. This simple-to-use, robust, semi-quantitative ELISA is a sensitive and specific method to aid in optimal process performance for purification, process control, routine quality control, and product release testing. This kit is “generic” in that it is designed to react with most HCPs that could contaminate the product regardless of the purification process. This Vero Cell 2G HCP ELISA Kit has a LOD of ~1.4 ng/mL. Use Sample Diluent, Item # I028, with these kits; available separately.

Product Information

Product Type: ELISA Kit

Target expression system: Vero Cell

Species group: mammal

Species: African green monkey

Assay format: 96-well plate

Time to result: ~2 hrs. 50 minutes

LOD: ~1.4ng/mL

LLOQ: ~3ng/mL

Recommended thinner: I028

Intended Use

This kit is designed to determine the presence of host cell protein impurities in products made by expression in Vero cells. The kit is for research and manufacturing use only and is not intended for diagnostic use in humans or animals.

Kit Contents

• Anti-Vero Cell 2G: HRP Affinity purified goat antibody conjugated to HRP in a preserved protein matrix. 1x12mL
• Anti-Vero Cell 2G Microtiter Strips 12×8-well strips in a bag with desiccant
• Vero Cell HCP Standards 2G Vero cell HCP solubilized in preserved bovine serum albumin. Standards at 0, 3, 6, 12, 25, 50, 100 and 200ng/mL. 1ml/vial
• 0.5M sulfuric acid stop solution. 1x12mL
• TMB Substrate 3,3′,5,5′ Tetramethylbenzidine. 1x12mL
• Concentrated wash (20X) Tris-buffered saline with preservative. 1x50ml

Storage

1. All reagents should be stored at 2-8°C to be stable until the expiration date printed on the kit.
2. After prolonged storage, you may notice a precipitate of salt and/or a yellowish colour of the wash concentrate. These changes will not affect the performance of the assay. To dissolve the precipitate, mix the wash concentrate well and dilute as directed in the ‘Reagent Preparation’ section.
3. Reconstituted Wash Solution is stable through the kit expiration date.

Performance Characteristics

This rating is generic in nature and is intended to supplement, but not replace, certain user and product-specific ratings and qualifications that should be made by each laboratory. At a minimum, each laboratory is encouraged to perform a peak and recovery study on their sample types.

Additionally, any of your sample types that contain process-derived HCPs within or above the analytical range of this essay should be evaluated for dilutional linearity to ensure the assay is accurate and has sufficient excess antibodies for your particular HCPs. Each laboratory and technician must also demonstrate their proficiency in the assay by conducting a precision study similar to the one described below.

Sensitivity

• The lower limit of detection (LOD) is defined as the concentration corresponding to a signal three standard deviations above the mean of the zero standards. The LOD is ~1.4 ng/mL.
• The lower limit of quantification (LLOQ) is defined as the lowest concentration, where the concentration coefficients of variation (CV) are less than 20%. The LLOQ is ~3 ng/mL.