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QCdetect(TM) Residual DNA Detection Kit for CHO cells

for Genetic Research
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at -20 degrees C.
  • Structural Formula
  • Label
  • Packing
SDS
Comparison
Product Number
Package Size
Price
Inventory
Distributor
294-85201
Barcode No
4548995093219
100Tests
List Price
JPY 143,000

In stock in Japan

Document

SDS
Product Specification Sheet
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

Kit Contents

1 x PCR Master Mix 2 x 1 mL
DNA Dilution Buffer (DDB) 1 x 10 mL
CHO Control DNA 1 x 40 μL

Overview

This product is a kit for quantifying genomic DNA from CHO cells by qPCR (probe method). It can be used for residual DNA testing for antibody pharmaceuticals and their manufacturing processes.

  • Features

    • Limit of detection: ≥ 0.0003 pg/test
    • Limit of quantitation: ≥ 0.003 pg/test
    • Calibration curve with high linearity
    • Minimal inter-assay variability and high reproducibility
    • Easy operation using the pre-mix buffer
    • Less susceptible to contaminants in samples
    • Contains an internal control
  • Detection wavelength

    CHO gDNA 520nm (e.g., FAM)
    Internal Control 554nm (e.g., HEX)

    Note) Internal Control is non-natural synthetic DNA.

Application

Calibration curve

qPCR of 0.0003 pg to 30,000 pg of CHO gDNA was performed using this kit to prepare a calibration curve (n = 3).

A calibration curve with very high linearity was obtained over a wide concentration range from 0.0003 pg to 30,000 pg.

 

294-85201_img03png.png
Pg/well 0.0003 0.003 0.03 0.3 3 30 300 3000 30000
Ct 37.639 34.822 31.546 28.117 24.809 21.337 18.079 14.726 11.506
37.107 34.981 31.506 28.324 24.906 21.416 18.084 14.744 11.467
37.642 34.689 31.595 28.267 24.987 21.533 18.225 14.807 11.269
Average Ct 37.462 34.830 31.549 28.236 24.901 21.428 18.129 14.759 11.414
S. E 0.30806 0.14652 0.04455 0.10713 0.08904 0.09853 0.0831 0.0424 0.12685

 

Detection of fragmented DNA

CHO gDNA was fragmented by sonication and detected by this kit to examine whether the fragmented CHO gDNA could be detected.

Fragmented gDNA could be detected with the same sensitivity as intact gDNA. A decrease in detection sensitivity was not observed, even for low concentrations of gDNA.

294-85201_img02png.png

 

Example of use for a high-protein sample -combined with DNA Extractor® Kit-

CHO gDNA was spiked to a sample containing high concentration of γ-globulin, and DNA was extracted by DNA Extractor® Kit.
The obtained DNA was quantified with QCdetect™ Residual DNA Detection Kit for CHO cells, and the spike recovery rate was obtained.

CHO gDNA could be recovered with a high recovery rate, even when a high-protein sample was used.

  • Sample composition

    • 20 mg/mL γ-globulin
    • 3% Mannitol
    • 2% Sucrose
    • 10 mM L-arginine
    • 0.01% Tween20

    Concentration of spiked CHO gDNA

    10 ng/mL
    1 ng/mL
    100 pg/mL

  • 04439536_img06.png

Detection Wavelength

CHO gDNA 520nm (e.g. FAM)
Internal Control 554nm (e.g. HEX)

Internal Control is non-natural synthetic DNA.

Overview / Applications

Outline This kit detects host cell-derived genomic DNA remaining in biological products such as vaccines and biopharmaceuticals by the qPCR methods using the TaqMan probe.
Our unique polymerase configuration enable highly sensitive detection of trace amounts of DNA contained in samples.

In order to suppress human error during liquid preparation and improve the work efficiency and the test accuracy, the PCR reaction solution is prepared in one bottle as 1 x PCR Master Mix.
In addition, it contains the primers/probes and the templates for internal control, so you can confirm that the PCR reaction is performed correctly.

When used in combination with the separately sold [DNA Extractor Kit], it is possible to consistently extract and detect genomic DNA in protein solution prepared using CHO cells.
Purpose Detection of residual DNA derived from the host cells, which is one of the quality control for biopharmaceuticals.
Application [Preparation of CHO control DNA for standard]
1) Mix 10uL of CHO Control DNA included in the kit and 990uL of DNA Dilution Buffer included in the kit.
2) Dilute the CHO Control DNA diluted in step 1) 10-fold with DNA Dilution Buffer to prepare a dilution series of 0.003pg/uL to 300pg/uL.
3) Perform the PCR reaction according to the package insert.

Property

Specificity Confirmed that amplification does not occur in human, E. coli, and yeast DNA.

Manufacturer Information

Alias

For research use or further manufacturing use only. Not for use in diagnostic procedures.

Product content may differ from the actual image due to minor specification changes etc.

If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.

The prices are list prices in Japan.Please contact your local distributor for your retail price in your region.

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