| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Storage |
Description
CHO Host Cell DNA Residue Detection Kit is used for the quantitative analysis of CHO host cell DNA residuce in intermediate samples, semi-finished and finished products of various biological products.
This kit adopts Taqman fluorescent probe and the polymerase chain reaction (PCR) method, which has fg level minimum detection limit and can specifically and quickly detect the residual CHO cell DNA. The kit needs to be used together with the the Residual DNA Sample Preparation Kit (Cat# 18461ES or 18469).
Features
High anti-interference capability – In complex sample matrices, spike recovery rates are consistently between 70% and 130%.
High sensitivity – Limit of quantification (LLOQ) as low as 10 fg/μL.
Excellent precision – Intra-assay CV <10%; inter-assay (intermediate) CV <15%.
Strong specificity – Specifically detects HEK 293 residual DNA without interference from other genomic DNA.
Robust contamination control – Includes UDG enzyme to digest potential carryover from PCR products and aerosols, preventing non-specific amplification.
Comprehensive validation – Fully validated according to ICH Q2 (R2) guidelines; complete validation reports available.
Regulatory compliance – Verified according to EP 2.6.7, JP G3, and USP <63> pharmacopeia standards.
Quality assurance – All enzymes are self-developed and industrialized, ensuring stable supply and consistent kit performance.
Components
|
Components No. |
Name |
41331ES50 (50 T) |
41331ES60 (100 T) |
|
A |
HEK293 qPCR Mix (UDG plus) |
0.75 mL |
1.5 mL |
|
B |
HEK293 Primer&Probe Mix |
250 μL |
500 μL |
|
C |
DNA Dilution Buffer |
2×1.8 mL |
4×1.8 mL |
|
D |
HEK293 DNA Control(30 ng/μL) |
25 μL |
50 μL |
Specifications
|
Linearity |
Standard curve range: 30 fg/μL – 300 pg/μL Correlation coefficient (R²): ≥ 0.98 Amplification efficiency: 90%–110% |
|
Accuracy |
Recovery rate: 50%–150% (industry standard 70%–130%) |
|
Precision |
Repeatability: CV < 15% Intermediate precision: CV < 15% |
|
Sensitivity |
Limit of quantification (LOQ): 10 fg/μL |
|
Specificity |
No interference from other genomic DNA sources |
|
Stability |
Accelerated stability: 14 days at 37 °C Freeze–thaw stability: ≥ 10 cycles |
|
Blank Control |
No-template control (NTC): In 96 repeated qPCR runs, all Ct values ≥ 35 or no amplification observed |
|
Applicable instrument models |
Bio-Rad: CFX96 Optic Module. Thermo Scientific: ABI 7500; ABI Quant Studio 5. SLAN-96S. |
|
Application |
Detection of residual HEK293 host cell DNA in viral and cell products, including lentivirus and AAV production, as well as other HEK293-derived bioproducts. |
Storage
This product should be stored at -25~-15℃ for 2 years.
Both 41331-A and 41331-B should be stored protected from light.
Figures
1. Good Linearity
Figure 1. Standard curve (left) and amplification curve plot (right) for HEK293 DNA (3G).
The linear range of the HEK293 Host Cell Residual DNA Detection Kit (3G) is 30 fg/μL to 300 pg/μL, with an R² value of 1.0 and an amplification efficiency of 100.45%. The coefficient of variation (CV) for detection values across concentrations is <15%.
2. Sensitivity: Limit of quantification (LOQ) - 0.3 fg/μL
Figure 2. qPCR detection of HEK293 DNA at 10 fg/μL.
HEK293 DNA (3G) at the lowest standard curve concentration (3 fg/μL) and below was tested with 10 replicates per concentration. Results showed that at concentrations ≥10 fg/μL, the CV was <20%, indicating that the limit of quantification (LOQ) of the HEK293 Host Cell DNA Residual Detection Kit (3G) is 10 fg/μL.
3. High Recovery rate: 50%–150%
Table 1. Recovery of HEK293 DNA in Blank Samples
Spike recovery rates for all concentrations tested were between 70% and 130%, with coefficients of variation (CV) below 20%, demonstrating consistent performance of the assay across a wide dynamic range.
Table 2. HEK293 DNA Recovery in Various Sample Matrices
HEK293 DNA (3 pg/μL) was spiked into five different background matrices (high protein, high nucleic acid, high salt, high pH, low pH) and extracted using the same procedure. Recovery rates ranged from 70% to 130%, with CVs below 20% for all matrices, demonstrating robust performance across diverse sample backgrounds.
Documents
Safety Data Sheet
Manuals
Host cell DNA (HCD) residue testing is required for biopharmaceutical manufacturing under ICH Q5A, ICH Q6B, FDA 21 CFR Part 610, and EMA guidelines. Acceptable residual HCD limits are typically ≤10 ng/dose (WHO guideline) or ≤100 pg/dose for continuous cell line-derived products, depending on the product class and route of administration. This kit provides qPCR-based quantification traceable to validated reference standards.
This kit uses primers and probes targeting genome-specific repetitive elements (e.g., Alu elements for human HEK293/CHO, LINE-1 for mammalian cells, or species-specific rDNA repeats for yeast/insect cell lines) that are highly conserved and abundant in the host genome. This provides high sensitivity — typically detecting sub-picogram quantities of residual host DNA — without cross-reacting with the target product or plasmid sequences.
Yes. The quantitative qPCR format of this kit is suitable for lot release testing, process characterization, and stability monitoring of biopharmaceutical drug substances and drug products. To generate regulatory-compliant data, assay performance qualification (sensitivity, specificity, linearity, spike recovery) should be documented per applicable guidelines. Contact your QA team to confirm kit suitability for your specific regulatory submission.
The LOD varies by kit generation and target cell line (e.g., HEK293, CHO, E. coli, Pichia pastoris). Third-generation (3G) kits typically achieve LOD values of 0.1–1 pg of host cell DNA per reaction. The validated linear range spans 4–6 orders of magnitude, supporting both low-level process monitoring and high-level spiking experiments. Validated LOD and LOQ values are provided in the product-specific datasheet.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
