HCP ELISA Kit
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- Target See all HCP products
- HCP (Host Cell Proteins (HCP))
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Reactivity
- Escherichia coli
- Detection Method
- Colorimetric
- Method Type
- Sandwich ELISA
- Detection Range
- 2.6 ng/mL – 250.0 ng/mL
- Minimum Detection Limit
- 2.6 ng/mL
- Application
- ELISA
- Purpose
- AccuSignal™ E. coli HCP ELISA Kit
- Brand
- AccuSignal™
- Analytical Method
- Quantitative
- Specificity
- Escherichia coli (E. coli) is a widely utilized organism in scientific research and biomanufacturing of therapeutic biologic drug substances. Monitoring the concentrations of impurities, particularly E. coli Host Cell Proteins (HCP), is of paramount importance in biomanufacturing processes. The primary objective of purification processes is to eliminate HCPs to minimize immunogenicity, enhance potency, and ensure the stability of a pure drug substance.
- Characteristics
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Synonyms: host cell proteins, ELISA, process development
Background: The antibodies in the kit have been specifically designed to detect HCP contaminants and to measure analytes from crude cellular harvests to process intermediates. The E. coli ELISA Kit uses purified antibodies generated against a mixture of proteins of mock induced E. coli cell lysate. These antibodies react strongly with E. coli cell lysates from a commonly used E.coli strain, BL21. The key component of this kit is a broad coverage antibody produced against a spectrum of host cell proteins including both low abundancy and high abundancy proteins.
- Components
- This kit contains: E. coli-HCP antibody coated 96-well strip plate, Plate Sealer, E. coli-HCP standard, E. coli-HCP detection antibody, sample buffer, Wash Buffer, TMB Buffer, Stop Buffer, and protocol.
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- Application Notes
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Application Note:The AccuSignal™ E. coli HCP ELISA Kit is a sandwich, enzyme-linked immunosorbent assay. The impurities in process-derived samples containing E. coli proteins are captured and detected in a microtiter-type plate by using pre-coated, purified, rabbit polyclonal, E. coli-HCP-specific antibodies. The immunological reaction after the addition of TMB results in a chromogenic change that can be measured photometrically at 450 nm with reference wavelength at 630–650 nm. The O.D. in the wells is directly proportional to the amount of HCP concentration and correlates to the amount of HCP present in the original sample. The HCP concentration can be calculated based on the calibration curve generated with the provided E. coli-HCP standard.
- Plate
- Pre-coated
- Protocol
- E. coli HCPs from a sample are captured on the AccuSignal™ E. coli HCP ELISA Kit plate, which contains pre-immobilized anti-E. coli HCP antibodies. The kit detection antibodies (anti-E. coli HCP antibodies) are conjugated with Horse Radish Peroxidase (HRP). Upon exposure to 3,3’,5,5’-Tetramethylbenzidine (TMB) substrate, HRP activates, resulting in a color change that can be measured with a plate reader at 450 nm. This optical density (OD) value is directly proportional to the concentration of E. coli HCPs, which can be calculated by interpolating against a Four-Parameter Logistic (4PL) standard calibration curve.
- Assay Precision
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Precision (Intra-Assay Variability): 20%
Precision (Inter-Assay Variability): 25% - Restrictions
- For Research Use only
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- Storage
- RT,4 °C,-20 °C
- Storage Comment
- The E. coli HCP Kit and reagents should be stored at 2-8°C. Allow reagents to reach room temperature (18-26°C) before use and may be used until the expiration date. It is recommended to aliquot the reconstituted standard
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- Target See all HCP products
- HCP (Host Cell Proteins (HCP))
- Alternative Name
- Host Cell Proteins (HCP Products)
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