| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Storage |
Description
Sf9 and Baculovirus DNA Residue Detection Kit is used for quantitatively analysing and detecting Sf9 host cell DNA and baculovirus DNA residues in the development and production of a variety of biologics, such as gene therapies, recombinant proteins and vaccines, using the baculovirus-insect cell (Sf9) expression system.
This kit is based on the principle of fluorescence quantitative PCR by probe method, and uses multiplex qPCR method to detect Sf9 and baculovirus residual DNA, respectively. The limit of quantification can be up to 1 fg/μL for Sf9, and the limit of quantification for baculovirus can be up to 7 copies/uL.The kit needs to be used together with the the Residual DNA Sample Preparation Kit (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 lower limit of quantification (LLOQ) was 1fg/μL for Sf9 and 7copies/uL for AcNPV;High precision: high intra-batch reproducibility CV <10%, small inter-batch variation i.e. intermediate precision CV <15%;
- Strong specificity: specific detection of Sf9 and AcNPV residual DNA without interference from other exogenous genomic DNA;
- Strong anti-interference: the introduction of internal quality control (IC) can exclude sample interference, abnormal reaction preparation and other factors, effectively avoiding false negatives.
Application
Residual Sf9 and Baculovirus DNA test in biological products.
Specification
|
Sensitivity |
Sf9 1fg/μL，AcNPV 7copies/uL (range （Sf9 3fg/μL~300pg/μL，AcNPV 2.12×101～2.12×106copies/μL）) |
|
Assay Time |
~1.5 hours |
|
Assay Principle |
Fluorescent probe qPCR method |
|
Required Instruments |
Real-time fluorescence quantitative PCR system |
Components
|
Components No. |
Name |
41330ES50-EN |
41330ES60-EN |
|
41330-A |
Sf9&Baculovirus qPCR Mix |
0.75 mL |
1.5 mL |
|
41330-B |
Sf9&Baculovirus Primer&Probe Mix |
200 μL |
400 μL |
|
41330-C |
DNA Dilution Buffer |
1.8 mL×2 |
1.8 mL×4 |
|
41330-D |
Sf9&Baculovirus DNA Control Mix(30 ng/μL & 2.12×109 copies/μL) |
25 μL |
50 μL |
|
41330-E |
IC* |
50 μL |
100 μL |
*IC：Internal control.
Storage
This product should be stored at -25~-15℃ for 2 years.
Both 41330-A and 41330-B should be stored protected from light.
Figures
Limit of quantification
Sf9 DNA at concentrations of 3 fg/μL, 1 fg/μL, and 0.5 fg/μL, and baculovirus AcNPV DNA at concentrations of 2.12×101copies/uL, 7copies/uL, and 3.5copies/uL were detected, with 10 replicates for each concentration, respectively. The results showed that the CV of Sf9 DNA was <20% at concentrations of 1 fg/μL and above, and the CV of baculovirus AcNPV DNA was <20% at concentrations of 7copies/uL and above. Therefore, the limit of quantification in the Sf9 and baculovirus AcNPV DNA residue detection kits was 1 fg/μL for Sf9 and 7copies/uL for baculovirus AcNPV.
|
Test item |
Sf9 DNA (fg/uL) |
AcNPV (copies/μL) |
||
|
Quantity |
Return ratio |
Quantity |
Return ratio |
|
|
1 |
0.81 |
81% |
5.11 |
73.00% |
|
2 |
0.84 |
84% |
5.22 |
74.57% |
|
3 |
0.91 |
91% |
8.48 |
121.14% |
|
4 |
0.92 |
92% |
7.04 |
100.57% |
|
5 |
0.78 |
78% |
7.21 |
103.00% |
|
6 |
1.12 |
112% |
6.76 |
96.57% |
|
7 |
0.71 |
71% |
6.24 |
89.14% |
|
8 |
1.02 |
102% |
7.06 |
100.86% |
|
9 |
1.06 |
106% |
6.62 |
94.57% |
|
10 |
0.78 |
78% |
6.64 |
94.86% |
|
Average value |
0.90 |
\ |
6.64 |
\ |
|
CV |
15.06% |
\ |
14.70% |
\ |
Table 1. Limit of quantification (LOQ) assay results for Sf9 and baculovirus AcNPV DNA
Documents：
Safety Data Sheet
Manuals
41330_Manual_Ver.EN20250901.pdf
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.
