Intra-assay: 4.2%;
Inter-assay: 5.7%
E. coli host cell proteins (HCP) constitute a diverse and functionally intricate group of macromolecules within the Escherichia coli cell. The host proteins of E. coli play crucial roles in various cellular processes. These proteins are involved in fundamental metabolic pathways such as glycolysis, the tricarboxylic acid cycle, and amino acid biosynthesis, which are essential for the survival and growth of the bacterium. Recombinant expression in E. coli is a simple and cost-effective method for producing certain proteins. These proteins, often used as therapeutic agents, must be highly purified. The manufacturing process may introduce contamination by HCPs from E. coli, which can reduce efficacy and cause adverse reactions. Therefore, minimizing HCP impurities is crucial. This simple, sensitive, and fast OneStep ELISA is a powerful method for optimizing purification processes, process control, routine quality control, and product release testing.
Standard curve
Example of E. coli Host Cell Proteins standard curve in Assay Diluent #8.
Linearity
The concentrations of E. coli Host Cell Proteins were measured and interpolated from the target standard curves and corrected for sample dilution.
Undiluted samples are as follows: E. coli expression purification sample (100%). The interpolated dilution factor corrected values are plotted. The mean target concentration was determined to be 19.18 ug/mg in E. coli expression purification sample.
Linearity
Undiluted samples are as follows: unpurified recombinant E. coli protein expressed in Luria-Bertani (LB) medium (100%). The interpolated dilution factor corrected values are plotted. The mean target concentration was determined to be 10.23 ng/mL in cell culture medium.
Specificity
Test the purified recombinant proteins from different hosts as follows: Yeast sample(1ug/mL); E. coli sample(1ug/mL); HEK 293 sample(1ug/mL).
Specificity
The concentrations of E coli host cell proteins were quantified prior to and following the HCP removal process. The sample is one of E. coli expression purification sample from Starter catalog protein (1ug/mL).
Specificity
Test the HCP standard protein as follows: E. coli HCP standard (25ng/mL); HEK293 HCP standard (200ng/mL); CHO HCP standard (200ng/mL).
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Protocol Diagram