Description

This kit utilizes a solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual host cell proteins (HCPs) from HEK293 cells in the sample. A polyclonal antibody specific to HEK293 HCPs was employed in the assay to capture any remaining HCPs in the sample. Both the Calibration standards (or test sample) and the HRP (Horseradish Peroxidase) labeled anti-HEK293 HCP antibody were simultaneously added to the microtiter plate, which coated with the affinity purified capture antibody and followed by incubation and washing. Then TMB (3,3',5,5' -tetramethylbenzidine) substrate was added into reaction, HRP catalyzed the oxidation of TMB by H₂O₂ to produce a blue 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 450nm). The absorbance values at450 nm wavelength were positively correlated with the HCPs concentration in the Calibration standard and the sample. The concentration of HCPs in the sample can be calculated using the dose-response curve.

Components	Shipping Condition	Unit Size	Detection Method
HEK293 HCP Calibration Standard	2-8℃	96 Tests	Sandwich ELISA
Anti-HEK293 HCP Microtiter Strips
Reconstitution Solution
Diluent
Wash Buffer Concentrate (10×)
Anti-HEK293:HRP (100×)
TMB Substrate
Stop Solution
Sealing Film

Specifications

Linearity & Range	6-540 ng/mL, R2>0.990
Accuracy	Recovery Rate= 96.7%-102.9%
LLOQ	6 ng/mL
LOD	1 ng/mL
Precision	2.4%-3.7% (CV≤20%)
Antibody Coverage (IMBS-2D)	67.1%-82.8%
Antibody Coverage (IMBS-LC/MS)	79.70%
Specificity	No cross-reactivity with P.pastoris and Sf9 cell.
Robustness	Incubate at 25±3℃, CV≤ 20%
	Incubation speed: 500-600 rpm
	Instrument suitability not limited to Thermo Multiskan FC, MD Spectra Max ABS Microplate Readers

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Documents

User Guide

HEK293 HCP ELISA Kit SHENTEK®