| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
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Description
The Vero Host Cell Residue DNA Size Analysis Kit is designed for the quantitative analysis of Vero residual DNA fragments of various lengths in intermediates, semi-finished products, and final products of biological preparations.
This kit utilizes the qPCR fluorescence probe principle to specifically and rapidly detect Vero DNA residues both below and above 200 base pairs, with a quantification limit as low as 10 fg/μL. It also includes the Vero DNA Control (DNA quantitative reference). This kit can be used in conjunction with the company's magnetic bead-based residual DNA sample preparation kits (Cat#18461ES).
Feature
- Compliance with regulations: the products are fully validated according to the requirements of Chp, USP, ICH, etc., and their performance is in line with Chinese and foreign regulations and standards;
- Guarantee of quality: the raw materials of the kits are all self-developed, and the qPCR Mix and other enzyme products are produced in an ultra-clean enzyme factory;
- High sensitivity: the limit of quantification can reach 3fg/μL level;
- High precision: high intra-batch repeatability and low inter-batch variation;
- Strong exclusivity: specific detection of residual DNA in Vero cells without interference from other exogenous genomic DNA;
- Strong anti-interference: add internal control (IC), easy to exclude sample interference, reaction preparation abnormalities and other factors.
Application
Residual Vero Host Cell DNA Size test in biological products
Specification
|
Sensitivity |
3fg/μL (range 300pg/μL~3fg/μL) |
|
Assay Time |
~1.5 hours |
|
Assay Principle |
Fluorescent probe qPCR method |
|
Required Instruments |
Real-time fluorescence quantitative PCR system |
Components
|
Components No. |
Name |
41314ES70-EN |
41314ES74-EN |
|
41314-A |
Vero qPCR Mix |
0.75 mL×4 |
1.5 mL×4 |
|
41314-B1 |
Vero Primer&Probe Mix-97 |
200 μL |
400 μL |
|
41314-B2 |
Vero Primer&Probe Mix-154 |
200 μL |
400 μL |
|
41314-B3 |
Vero Primer&Probe Mix-219 |
200 μL |
400 μL |
|
41314-B4 |
Vero Primer&Probe Mix-507 |
200 μL |
400 μL |
|
41314-C |
DNA Dilution Buffer |
1.8 mL×2 |
1.8 mL×4 |
|
41314-D |
Vero DNA Control (30 ng/μL) |
25 μL |
50 μL |
|
41314-E |
IC |
200 μL |
400 μL |
Storage
This product should be stored at -25~-15℃ for 2 years.
Both 41314-A and 41314-B1&B2&B3&B4 all should be stored protected from light.
Figures
- Limit of quantification
The primer probes of the four fragments, 97bp, 154bp, 219bp and 507bp, were used to detect Vero DNA at 30pg/μL, 300fg/μL, and 3fg/μL, respectively, with 10 replicates for each concentration. The results showed that at concentrations of 3 fg/μL and above, the CVs of the four fragments were <20%, and the backcalculated ratios were between 70% and 130%, so the limit of quantification of the four fragments of the Vero Residual DNA Fragmentation Assay Kit was 3 fg/μL.
|
Repetition number
Test item |
Vero DNA (fg/uL) |
|||||||
|
97bp |
154bp |
219bp |
507bp |
|||||
|
Quantity |
Return ratio |
Quantity |
Return ratio |
Quantity |
Return ratio |
Quantity |
Return ratio |
|
|
1 |
3.49 |
116.33 |
2.48 |
82.67 |
3.33 |
111.00 |
3.53 |
117.61 |
|
2 |
3.03 |
101.00 |
3.23 |
107.67 |
2.62 |
87.33 |
3.56 |
118.65 |
|
3 |
3.64 |
121.33 |
3.10 |
103.33 |
3.52 |
117.33 |
3.34 |
111.50 |
|
4 |
3.01 |
100.33 |
2.55 |
85.00 |
3.58 |
119.33 |
3.36 |
112.03 |
|
5 |
2.92 |
97.33 |
3.15 |
105.00 |
3.53 |
117.67 |
2.66 |
88.64 |
|
6 |
2.96 |
98.67 |
2.77 |
92.33 |
3.34 |
111.33 |
2.93 |
97.80 |
|
7 |
3.33 |
111.00 |
2.88 |
96.00 |
3.14 |
104.67 |
3.35 |
111.52 |
|
8 |
2.34 |
78.00 |
3.32 |
110.67 |
3.01 |
100.33 |
2.69 |
89.79 |
|
9 |
2.84 |
94.67 |
3.84 |
128.00 |
2.85 |
95.00 |
3.33 |
111.01 |
|
10 |
2.89 |
96.33 |
2.88 |
96.00 |
3.00 |
100.00 |
3.47 |
115.67 |
|
Average value |
3.05 |
\ |
3.02 |
\ |
3.19 |
\ |
3.22 |
\ |
|
CV |
12.09% |
\ |
13.26% |
\ |
10.07% |
\ |
10.39% |
\ |
Table 1. Detection results of limit of quantification for four fragments of 97bp, 154bp, 219bp and 507bp
Documents:
—
What regulatory guidelines govern host cell DNA residue testing?
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.
How does this kit achieve host cell-specific detection?
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.
Can this kit be used for lot release or stability studies?
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.
What is the limit of detection (LOD) of this host cell DNA detection kit?
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.
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