Description

This e coli hcp assay kit is designed to quantify and characterize host cell proteins (HCPs) in products manufactured by expression in E.coli originated from BL21, such as interleukin (IL), recombinant human interferon (rhIFN), recombinant human tumor necrosis factor (rhTNF), growth-promoting factor (EGF/FGF/PDGF), et.al.

This kit utilizes a solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual E.coli (Protein expression strains) host cell proteins (HCPs) in the samples. Polyclonal antibody specific to E.coli (Protein expression strains) HCPs was employed in the assay to capture any remaining HCPs in the samples. Both the Calibration Standard and test samples were simultaneously added to the microtiter plate coated with affinity purified capture antibody, and followed by incubation and washing. The biotinylated antibody was added to the microtiter plate to bind the HCPs, then reacted with streptavidin labeled with HRP (Horseradish Peroxidase). TMB (3,3',5,5' -tetramethylbenzidine) substrate was added into reaction, HRP catalyzed the oxidation of TMB by H₂O₂ to produce a blue colored product (maximum absorption peak at 655 nm). Then the stop solution was added to terminate the enzymatic reaction, resulting in a yellow colored product (maximum absorption peak at 450 nm). The absorbance values at 450 nm wavelength were positively correlated with the HCPs concentration in the calibration Standard and the samples. The concentration of HCPs in the sample can be calculated using the dose-response curve.

Components	Shipping Condition	Unit Size	Detection Method
E.coli HCP-E Calibration Standard	2-8℃	96 Tests	Sandwich ELISA
Anti-E.coli HCP-E Microtiter Strips
Reconstitution Solution
Diluent
Wash Buffer Concentrate (10×)
Anti-E.coli HCP-E: Biotinylated Conjugate (100×)
Streptavidin-HRP (100×)
TMB Substrate
Stop Solution
Sealing Film

Specifications

Linearity & Range	1-243 ng/mL, R²>0.990
Accuracy	Recovery Rate= 87.1%-108.3%
LLOQ	1.5 ng/mL
LOD	0.0113 ng/mL
Repeatability	3.5%-4.9% (CV≤20%)
Antibody Coverage (IMBS-2D)	70.6%-100%
Antibody Coverage (IMBS-LC/MS)	88.80%
Specificity	No cross-reactivity with CHO, Vero, HEK293T, Hansenula polymorpha HCPs
Robustness	Incubate at 25±3℃, CV≤ 20%
Robustness	Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2

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Documents

User Guide

E.coli (Protein Expression Strains) HCP ELISA Kit SHENTEK®