CHO HCP (Host Cell Protein) ELISA Assay
The CHO HCP (Host Cell Protein) ELISA Assay is For Research Use Only
Size: 1×96 wells
Sensitivity: 5 ng/mL
Calibrator Range: 6.25 ng/ml – 400 ng/ml
Sample Size: 100 uL
Sample Type: Cell Culture Supernatant and other Biological Preparations
Incubations Time: 2 hours and 30 minutes
Assay Background
A variety of proteins which are used as therapeutic agents in humans and animals are produced through recombinant expression in Chinese Hamster Ovary (CHO) cells. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from CHO cells, 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 CHO HCPs are reacted with already coated affinity purified capture anti-CHO HCP antibody. Following an incubation period unbound components are removed by a washing step. Then the anti-CHO HCP: HRP is added. This immunological reaction results in formation of a sandwich complex of solid phase antibody-HCP-enzyme labeled antibody. The wells are again washed to remove any unbound reactants. The TMB substrate is then added to the well. The amount of hydrolyzed substrate is read on a microtiter plate reader and it is directly proportional to the concentration of CHO HCPs present.
Reagent Preparation
- Bring all kit components and samples to room temperature (18-25°C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
- To make Wash Buffer (1X); dilute 25 ml of 20X Wash Buffer in 475 ml of DI water.
- Standard: Reconstitute the lyophilized standard in 1000ul of Standard diluent to get a concentration of 400ng/ml. Keep the standard for 15 minutes. 400ng/ml is the top standard. Perform serial dilution to get the remaining standards. Standard range for Octreotide ELISA is 400 ng/ml, 200 ng/ml, 100 ng/ml, 50 ng/ml, 25 ng/ml, 12.5 ng/ml and 6.25 ng/ml. Standard Diluent (1X) serves as the zero standard (0 ng/ml).
- After reconstitution, immediately use the standards and balance reconstituted standard should be discarded.
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