Host Cell Residual DNA Detection Aids Biologics R&D and Manufacturing

Host cell residual DNA is a process-related impurity involved in the production of biologics, which not only reduces the effectiveness of biologics, but also may pose safety concerns such as infectiousness or tumorigenicity. Therefore, regulatory agencies in various countries have set limits on the amount of residual DNA in biologics.

The current guidelines of WHO and US FDA recommend that residual DNA in finished products should not be higher than 10 ng/dose, and US FDA also points out that residual DNA in host cells of biological products should not be higher than 100 pg/dose. The General Principles of the European Pharmacopoeia stipulate that the limit of residual DNA in biological products should not be higher than 10 ng/dose, but the residual DNA limit for individual vaccines is more stringent, e.g., the residual DNA in the inactivated vaccine against hepatitis A should not be higher than 100 pg/dose, and that in the vaccine against hepatitis B should not be higher than 10 pg/dose. The 2020 edition of the Chinese Pharmacopoeia, Part III, stipulates that the DNA residue in biological preparations produced on a cellular matrix should not exceed 100 pg/dose, and the DNA residue in vaccines produced on a bacterial or fungal matrix should not exceed 10 ng/dose.

In addition, for the methods of exogenous DNA residue determination, national pharmacopoeias also give guidance recommendations. The U.S. Pharmacopoeia 2017 edition of USP40-NF35 General Provision 1130 describes three methods for the determination of exogenous DNA residues, which are DNA probe hybridization, threshold method and real-time quantitative PCR method. The European Pharmacopoeia proposes real-time quantitative PCR and immunoenzymatic method, two sensitive analytical methods for quantifying host cell residual DNA. The Chinese Pharmacopoeia 2020 edition of the three general rules 3407 also stipulates that the host cell DNA residue detection methods are DNA probe hybridization, fluorescence staining and quantitative PCR.

Among them, qPCR method has very high sensitivity, sequence specificity and accuracy, which can provide a reliable detection means for the biopharmaceutical industry in process research and quality control of finished products, and has now become the preferred detection method for each biological product manufacturer.

Yeasen Biotechnology has independently developed 4 qPCR kits for CHO&HEK293&Vero&E.coli, which can rapidly and efficiently detect host DNA residues in biopharmaceuticals.

 

 

Product Features

High Sensitivity：The lower limit of detection can be as low as 0.5 fg/μL.

High Precision: Repeatability CV <10%, Intermediate precision <15%.

Strong Proprietary：No cross-reactivity between different cell lines.

Quantitative Accuracy: Exclusive sample prep kit for magnetic bead assay.

Short Operating Time: Complete testing process within 3 hours.

 

Product Parameters

Product No.	41305ES	41306ES	41307ES	41308
Product name	CHO Residual DNA Detection Kit	HEK293 Residual DNA Detection Kit	Vero Residual DNA Detection Kit	E.coli Residual DNA Detection Kit
Product Component	CHO DNA Control CHO qPCR mix DNA Dilution Buffer	HEK293 DNA Control HEK293 qPCR mix (UDG plus)  HEK293 Primer&probe mix DNA Dilution Buffer	Vero  DNA Control Vero qPCR mix  Vero Primer&probe mix DNA Dilution Buffer	E.coli  DNA Control E.coli qPCR mix E.coli Primer&probe mix DNA Dilution Buffer
Product Specification	100T	100T	100T	100T
Appliance	Quantitative analysis of host cell DNA residues in intermediate, semi-finished and finished products of various biological products
limit of quantification	0.5 fg/μL	10fg/μL	0.5fg/μL	30 fg/μL
Pre-treatment kit	Magnetic Bead DNA Residue Pre-treatment Kit: Manual Extraction, Cat#18461ES, Adapts to All Residue Kits
Applicable models	Thermo：ABI 7500；ABI QuantStudio 5；ABI StepOnePlus； Bio-Rad：CFX96 Optic Module.

 

Workflow

Product Data

Residual DNA Detection Kit

Figure 1 Standard curve generated from a 10-fold dilution series of CHO standard DNA.

The results show that the host cell residual DNA quantitation kits exhibit a broad dynamic range and high sensitivity.

 

Magnetic Bead Method Sample Pre-treatment Kit

Table 1 DNA Recovery Using the MolPure® Magnetic Residual DNA Sample Preparation Kit performance data from independent validation study using E。coli genomic DNA spike per sample.

Sample	Concentration	Mean recovery	CV
300pg/uL	288.49pg/uL	96.16%	7.66%
150pg/uL	162.51pg/uL	108.34%	10.02%
10pg/uL	10.48pg/uL	104.89%	8.15%
5pg/uL	4.66pg/uL	93.20%	9.67%
60fg/uL	56.97fg/uL	94.95%	12.82%
30fg/uL	31.76fg/uL	104.91%	12.49%

 

Product Information

Description	Part Number
CHO Host Cell DNA Residue Detection Kit (2G)	41305ES
HEK293 Host Cell DNA Residue Detection Kit (2G)	41306ES
Vero Host Cell DNA Residue Detection Kit (2G)	41307ES
E.coli Host Cell DNA Residue Detection Kit (2G)	41408ES
MolPure® Magnetic Residual DNA Sample Preparation Kit	18461ES