| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Storage |
Description
Pichia pastoris Host Cell DNA Residue Detection Kit is used for the quantitative analysis of Pichia pastoris 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 Pichia pastoris cell DNA. 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 limit of quantification can reach 3 fg/μL level;
- High precision: high intra-batch repeatability and low inter-batch variation;
- Strong exclusivity: specific detection of residual DNA in Pichia pastoris 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 Pichia pastoris Host Cell DNA test in biological products.
Specification
|
Sensitivity |
3 fg/μL (range 30fg/μL~300pg/μL) |
|
Assay Time |
~1.5 hours |
|
Assay Principle |
Fluorescent probe qPCR method |
|
Required Instruments |
Real-time fluorescence quantitative PCR system |
Components
|
Components No. |
Name |
41328ES50-EN |
41328ES60-EN |
|
41328-A |
Pichia pastoris qPCR Mix |
0.75 mL |
1.5 mL |
|
41328-B |
Pichia pastoris Primer&Probe Mix |
200 μL |
400 μL |
|
41328-C |
DNA Dilution Buffer |
1.8 mL×2 |
1.8 mL×4 |
|
41328-D |
Pichia pastoris DNA Control (30 ng/μL) |
25 μL |
50 μL |
|
41328-E |
IC* |
50 μL |
100 μL |
*IC：Internal control.
Storage
This product should be stored at -25~-15℃ for 2 years.
Both 41328-A and 41328-B should be stored protected from light.
Figures
Limit of quantification
Pichia pastoris DNA was detected at 30 fg/uL, 10 fg/uL, 3 fg/uL, and 1 fg/uL with 10 replicates for each concentration. The results showed that at concentrations of 3 fg/uL and above, the CV was <20%, i.e., the limit of quantification of Pichia pastoris host cell DNA residues was 3 fg/uL.
|
Test item |
Pichia pastoris DNA (fg/uL) |
|
|
Quantity |
Return ratio |
|
|
1 |
3.83 |
127.76 |
|
2 |
3.68 |
122.53 |
|
3 |
2.55 |
85.05 |
|
4 |
2.66 |
88.63 |
|
5 |
3.74 |
124.63 |
|
6 |
3.29 |
109.54 |
|
7 |
3.04 |
101.42 |
|
8 |
3.54 |
118.1 |
|
9 |
3.09 |
102.91 |
|
10 |
3.27 |
109.01 |
|
Average value |
3.27 |
\ |
|
CV% |
13.49% |
\ |
Table 1. Limit of quantification (LOQ) assay results for Pichia pastoris DNA
Documents
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.
