E.coli HCP ELISA Assay Kit
The E.coli HCP ELISA Assay Kit is For Research Use Only
Size: 1 x 96 wells
Calibrator Range: 5 ng/ml – 100 ng/ml
Sensitivity: 5 ng/ mL
Sample Size: 50 uL
Sample Type: Cell Culture Supernatant and other Biological Preparations
Incubation Time: 4 hours and 30 minutes
Assay Background
A variety of proteins and pDNA which are used as therapeutic agents in humans and animals are produced through recombinant expression in E.coli. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from E.coli which may result in adverse toxic or immunological reactions that ultimately affect the efficacy of the therapeutic agent. The simple, objective and semi-quantitative ELISA is a highly sensitive method that aids in purification process development, process control, quality control and product release testing optimally.
Assay Principle
This assay is based on the Sandwich ELISA procedure. Samples containing E-coli HCPs are reacted with anti-E.coli:HRP antibody simultaneously in the microtiter wells already coated with affinity purified capture anti-E.coli antibody. This immunological reaction results in formation of a sandwich complex of solid phase antibody-HCP enzyme labeled antibody. The wells are washed to remove any unbound reactants as per the wash procedure (see Assay procedure section mentioned below). The substrate 3,3’,5,5’ Tetramethyl Benzidine is then reacted. The amount of hydrolyzed substrate is read on a microtiter plate reader and it is directly proportional to the concentration of E.coli HCPs present.
Handling & Storage
- All reagents should be stored at 20C to 80C for stability.
- All the reagents and wash solution are stable until the expiration date of the kit.
- Before using, bring all components to Room Temperature (18-25 ºC). Upon assay completion return all
components to appropriate storage conditions. - The Substrate is light-sensitive and should be protected from direct sunlight or UV sources.
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