| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Storage |
Description
E.coli Host Cell DNA Residue Detection Kit is used for the quantitative analysis of E.coli 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 E.coli cell DNA. The kit needs to be used together with the the Residual DNA Sample Preparation Kit (Cat# 18461ES).
Specifications
|
Cat.No. |
41308ES50-EN / 41308ES60-EN |
|
Size |
50 T-EN / 100 T-EN |
Components
|
Components No. |
Name |
41308ES50-EN |
41308ES60-EN |
|
41308-A |
E.coli qPCR Mix |
0.75 mL |
1.5 mL |
|
41308-B |
E.coli Primer&Probe Mix |
250 μL |
500 μL |
|
41308-C |
DNA Dilution Buffer |
2×1.8 mL |
4×1.8 mL |
|
41308-D |
E.coli DNA Control（30 ng/μL） |
25 μL |
50 μL |
Storage
This product should be stored at -25~-15℃ for 2 years.
Both 41308-A and 41308-B should be stored protected from light.
Applicable instrument models
Include but not limited to:
Bio-Rad: CFX96 Optic Module.
Thermo Scientific: ABI 7500; ABI Quant Studio 5.
Instructions
Figures
1.Good linearity: Linear range from 30 fg/μL ~ 300 pg/μL, R²= 1.000, Eff=101.74%, CV<15%.
Figure 1. Standard curve for E. coli DNA qPCR assay.
2. Specificity
To assess interference from host-cell genomic DNA used in biopharmaceutical production, genomic DNA from HEK293, Vero, and CHO cells was tested. No cross-reactivity or signal interference was observed, and the amplification curves fully overlapped with the negative controls.
Figure 2. Specificity/interference test results.
3. High Recovery
E. coli DNA (3 pg/μL) was spiked into five challenging matrices—high protein, high nucleic acid, high salt, high pH, and low pH. Each sample was extracted in duplicate and tested in triplicate by qPCR.
Across all matrices, recovery remained within 70–130% with CVs below 20%, confirming reliable performance under diverse sample conditions.
Table 1. Recovery of Spiked Simulated Samples
|
Sample Type |
Theoretical Concentration (pg/μL) |
Extraction 1 (pg/μL) |
Extraction 2 (pg/μL) |
Average Measured Concentration (pg/μL) |
CV (%) |
Recovery (%) |
|
Baseline Sample |
— |
— |
— |
— |
— |
— |
|
High Protein |
3 |
2.25 |
2.23 |
2.24 |
1.09 |
74.74 |
|
High Nucleic Acid |
3 |
3.93 |
3.96 |
3.95 |
1.56 |
131.52 |
|
High Salt |
3 |
2.69 |
2.67 |
2.68 |
2.63 |
89.20 |
|
High pH |
3 |
2.90 |
3.67 |
3.29 |
13.82 |
109.55 |
|
Low pH |
3 |
2.77 |
2.87 |
2.82 |
3.41 |
94.01 |
4. Limit of Quantification (LOQ): 30 fg/μL.
E. coli DNA at 50 fg/μL, 40 fg/μL, 30 fg/μL, 10 fg/μL, and 5 fg/μL was tested with 10 replicates per concentration. Results showed that at 30 fg/μL and above, the CV was < 20%, indicating that the limit of quantification (LOQ) of the E. coli Host Cell DNA Residual Detection Kit (2G) is 30 fg/μL.
|
Replicate |
E. coli DNA (fg/μL) |
Back-calculated (%) |
|
1 |
29.43 |
98.09 |
|
2 |
29.51 |
98.35 |
|
3 |
32.04 |
106.79 |
|
4 |
26.07 |
86.89 |
|
5 |
34.06 |
113.53 |
|
6 |
33.54 |
111.79 |
|
7 |
26.95 |
89.82 |
|
8 |
27.38 |
91.26 |
|
9 |
27.04 |
90.13 |
|
10 |
26.10 |
87.02 |
|
Mean |
29.21 |
— |
|
CV (%) |
10.40 |
— |
Figure 3. qPCR results for 30 fg/μL E. coli DNA
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.
