CHO PLBL2 ELISA
PLBL2 is recognized as a high-risk Host Cell Protein (HCP) due to its challenging removal during bioprocessing and its significant immunogenic potential. Despite sharing approximately 80% amino acid identity with its human ortholog, key structural differences make PLBL2 prone to triggering immune responses in patients.
Its clinical relevance is underscored by findings from Fischer et al. (2017), where approximately 90% of patients treated with lebrikizumab developed antibodies targeting PLBL2, demonstrating its role as a significant immunogenic impurity.
PLBL2 also presents unique challenges in purification workflows. It often co-purifies with therapeutic monoclonal antibodies (mAbs), likely due to interactions with the Fab´2 portion of the mAb, which can be influenced by the specific CHO cell clone used in production. Furthermore, its expression levels in CHO cells are heterogeneous, as highlighted by Vanderlaan et al. (2015), which can impact the performance of anti-CHO HCP ELISAs.
BioGenes PLBL2 ELISA Approach
BioGenes now offers a newly developed ELISA kit specifically designed for the detection of PLBL2. The antigen used in this kit is recombinant CHO PLBL2, expressed in HEK293 cells. By expressing the antigen in HEK cells, we eliminate potential cross-reactivity with other CHO HCPs.
For detection, we utilize polyclonal antibodies, specifically chosen for their superior versatility compared to monoclonal antibodies. Monoclonal antibodies, being epitope-specific, may fail to recognize native CHO PLBL2 if the protein structure differs from that of the PLBL2 antigen used for antibody generation. In contrast, polyclonal antibodies bind to multiple epitopes, providing a broader coverage and ensuring reliable detection of native Plbl2 protein. This comprehensive binding capability is shown by the exceptional assay sensitivity of the kit. The high sensitivity of PLBL2-HCP ELISA Kits is demonstrated by the validation parameters LOD and working range, as shown below:
- Limit of Detection (LOD): 0.015 ng/mL
- Working range: 0.078 - 5 ng/mL
In a comparative study against two competing products, the BioGenes kit demonstrated the highest sensitivity for detecting native CHO PLBL2 in real downstream processing (DSP) samples of CHO-expressed therapeutics. Its dynamic range ensures precise and reliable quantification, making it an ideal solution for monitoring these challenging HCPs in complex samples.
Your PLBL2 Comparison Study
PLBL2 ELISA Performance
The PLBL2-HCP ELISA method was qualified at BioGenes according to the respective ICH-Guideline ICH Q2(R1).