Zencore Analytical Sciences (AS) team has vast experiences in providing analytical support for biologics R&D, manufacturing, and the regulatory filings in FDA & China NMPA. Equipped with the state-of-the-art analytical tools, Zencore AS team can provide customers with analytical strategies and solutions for your products at the stage from pre-clinical to post approval changes.
A high-quality protein drug product should have stable physical and chemical properties, structural integrity, biological activities. Its stability that should meet the requirements from manufacturing, formulation process and the expected shelf life. The evaluation of this series of molecular quality attributes is called "developability" assessment.
The AS team can provide customers with testing services for the CQAs (critical quality attributes) and structural and functional characterization for a candidate drug during cell line development. The qualities attributes, which are normally monitored at this stage, include: size heterogeneity, charge variants, primary and higher order structure characterization, biological functional activity, and glycan profiles, etc.
The AS team can provide customers with services from method development to validation.
Which includes: impurity profiling, aggregation, charge variants characterization, glycan profiling, oxidation/deamidation/isomerization, disulfide bridge mapping, degradation pathway elucidation, and quantification of the process related impurities, etc.
Reference standard qualification.
Work with customers to set up specifications for DS and DP.
Perform the release testing of drug substances and drug products.
Conduct stability study.
Perform stress testing, accelerated stability, and long-term stability studies.
Validate and transfer analytical methods.
Head-to-head analysis of the batches before and after process changes.
Comparability study for IND or BLA filings.
Similarity assessment between an originator drug and the biosimilar products.
Comparability study required by the Prior Approval Supplement (PAS).
In use (aka compatibility) studies.
Leveraging our well-established analytical platform and cutting-edge technologies, the AS team is committed to providing efficient and high-quality services to our clients. Zencore AS team consists of seven groups (the biophysical, bioassay, mass spectrometry, physicochemical, ADC support, operation, and compliance team). A complete in-house analytical technology platform has been established and project specific analytical methods can be developed based on customers' requests.
Multiple binding assays have been developed for detecting the dual binding activities of bispecific antibodies. These assays can be utilized in the analysis of individual target binding, separately, or dual target binding, simultaneously. The binding affinity to each target is different for each arm of the bispecific antibody, this presents challenges for the development of a robust binding assay. The AS team investigated and optimized several conditions, such as the choice of coating antigen, antigen concentrations, incubation time and chromogenic reaction conditions, during the assay development. An ELISA method in the format of 1st antigen-antibody-biotinylated and 2nd antigen-enzyme labeled streptavidin was developed to detect dual target bindings simultaneously. All methods were fully validated and employed in the batch release and stability study.
Zencore AS team has developed a competitive ELISA for detecting fusion protein binding activity. The fusion protein has no Fc tag. A biotinylated fusion protein was first generated, then mixed with unlabeled fusion protein before mixing with the anti-fusion protein antibody in the binding assay. Then streptavidin labeled detection donor would bind to biotinylated fusion protein specifically, this would bring the donor closer to the fluorescent acceptor and complete the fluorescence resonance energy transferring (FRET). The FRET signal intensity is inversely related to the concentration of unlabeled protein sample. In this assay, homogeneous time-resolved fluorescence (HTRF) readout was achieved by FRET between donor of europium-XL665 complex and fluorescent acceptor. This method was validated with high sensitivity and stability.
For a peptide drug candidate, Zencore AS team generated the functional cell line by electroporating two vectors simultaneously, followed by clone screening and evaluation. The final cell line is capable of expressing the specific target protein which the peptide would specifically bind to and activate luciferase expression in the cells. The cell-based assay was validated and used in the batch release and stability study.
Zencore team has developed an analytical method for miRNA extraction, amplification and detection. Purified miRNA was transcribed to cDNA by reverse transcription, followed by a 3-step PCR to ligate the specified fragments and the amplification of the target sequence. Real-time fluorescence quantitative PCR was implemented to determine the concentration of the miRNA by analyzing the cycle threshold (Ct) value. Method validation is also completed in the follow-up study.
Accurate determination of sialic acid distribution (Z value) is of great significance for determining the quality attributes of the glycoprotein with high sialic acid content. ZhenGe applied the rapidly labeled 2-AB N-glycan testing kit (HiTang® N-sugar chain detection kit), which solved the systematic error caused by sialic acid shedding during sample pre-processing. This method enables accurate determination of the sialic acid distribution (Z-value).
