Ogataea polymorpha HCP ELISA
Ogataea polymorpha, together with Pichia pastoris and Saccharomyces cerevisiae, represents one of the three major yeast lineages, yet it shows markedly different cross-reactivity profiles. For certain therapeutics that are difficult to express, O. polymorpha offers significant advantages. Given that no dedicated HCP assay for O. polymorpha is currently available in the Chinese market, Cellgene Bioscience has developed a proprietary HCP ELISA kit specific to this host, providing robust support for drug development based on this expression system.
1.Background
1.1 Ogataea polymorpha Expression System
Ogataea polymorpha belongs to one of the major yeast lineages, and its taxonomic characteristics have been clearly described in the article ‘High-Specificity Yeast HCP ELISA: A Matrix Solution to Residual Detection Challenges’. Ogataea polymorpha is a representative strain of the Hansenula lineage. Beyond its applications in pharmaceutical engineering for the production of vaccines and enzymes, Ogataea polymorpha also plays an important role in functional food fermentation and chemical synthesis, including compounds such as paclitaxel and L-arabinose. Its key growth traits include methylotrophy, thermotolerance, osmotolerance, and the ability to oxidize hydrocarbons.
The functionality of Ogataea polymorpha is closely tied to the integrity of its eukaryotic organelles—such as mitochondria, nuclei, and peroxisomes—which provide specialized compartments essential for protein secretion, folding, and post-translational modifications (see Fig. 1)[1].
Figure 1
Electron Microscopy Image Abbreviations: N nucleus; M mitoehondrion; P peroxisome; V vacuole. Marker: 0.5 gin。
Given the advantages of the Ogataea polymorpha expression system in expression level, glycosylation capability, secretion efficiency, culture cost, scalability, applicable protein types, and glycoengineering complexity, the trend of developing biopharmaceuticals on this platform is steadily increasing. Approved vaccines and antibodies produced using this system include HPV vaccines (Cecolin, Cervarix), recombinant COVID-19 protein vaccine (ZF2001), and COVID-19 neutralizing antibodies (Amubarvimab/Romlusevimab), as summarized in Table 1. Consequently, there is a sustained and urgent market demand for HCP ELISA kits specific to Ogataea polymorpha.
Table 1
2.Quality Control Data：
Given the close phylogenetic relationship of Ogataea polymorpha with Saccharomyces cerevisiae and Pichia pastoris, we evaluated the cross-reactivity of the O. polymorpha HCP ELISA kit against HCPs from S. cerevisiae and P. pastoris (X-SS, GS115). The results showed minimal cross-reactivity, ranging from 0.4% to 3.3%, with both cases below 3.3%. The kit also demonstrated excellent background control, as detailed in the article “Background Reduction Strategy: Development of HCP ELISA.”
Additionally, multiple batch experiments were summarized to determine the linear range, R², and key QC parameters of the O. polymorpha HCP kit, including QL, DL, Recovery, CV, and others (see Tables 2-6 and Figures 2-5). The main features include, but are not limited to, high specificity, strong interference resistance, stable quantification limits, high precision, and clean background.
Table 2
2.1Cross-reactivity
Figure 2
Table 3
2.2 Interference Recovery
Table 4
2.3 LOD&LOQ
Figure 3
Table 5
2.4 Precision(CV)
Figure 4
Table 6
2.5 Ultra-low Blank
Figure 5
Reference Standard：
ICH Q6B: Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products.
ICH Q7: Good Manufacturing Practice Guide for Active Pharmaceutical Ingredients.
ICH S6 (R1).
EMA: Guideline on Immunogenicity assessment of therapeutic proteins (2017).
FDA: Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products (1997).
Reference：
[1] Klaas Nicolay, et al. A 31 P NMR study of the internal pH of yeast peroxisomes. Archives of Microbiology. 1987. 147(1):37-41.
Cellgene Bioscience has been dedicated to the biopharmaceutical and industrial testing field for 15 years, offering a series of HCP residual detection products, as well as comprehensive technical services including HCP-specific antibody development and coverage analysis.
https://www.elisakit.cc/
Cellgene Bioscience-Drug Residue Detection Products | |
Host Cell Protein ELISA kits (HCP) | |
CH-K0018-2 | CHO Host Cell Protein (CHO HCP) ELISA kit, G2 |
CH-K0018-3 | CHO Host Cell Protein (CHO HCP) ELISA kit, G3 |
HH-H0019-2 | HEK293 Host Cell Protein (HEK293 HCP) ELISA kit, G2 |
EH-E0020-3 | E. coli Host Cell Protein (E. coli HCP) ELISA kit, G3 |
PH-E0021-3 | Pichia pastoris Host Cell Protein (PP HCP) ELISA kit, G3 |
HP-H0023-3 | Ogataea polymorpha Host Cell Protein ELISA kit, G3 |
SC-H0024-3 | Saccharomyces cerevisiae Host Cell Protein ELISA kit, G3 |
SF-H0025-3 | Spodoptera frugiperda (Sf9) Host Cell Protein ELISA kit, G3 |
Medium Residues Detection kits | |
NEGES0890 | Protein A ELISA Kit (Boiling) |
NEGEP0890 | Protein A ELISA Kit |
NE03I0431 | Mouse Immunoglobulin G ELISA Kit |
NE11I0431 | Bovine Immunoglobulin G ELISA Kit |
NE01I0431 | Human Immunoglobulin G ELISA Kit |
NE06I0431 | Goat Immunoglobulin G ELISA Kit |
NEGEK0006 | Kanamycin ELISA Kit |
NEGEP1270 | Protein L ELISA Kit |
NEGEP1271 | Protein G ELISA Kit |
NEGES0014 | Bovine Serum Albumin ELISA Kit |
NEGES0015 | Human Serum Albumin ELISA Kit |
NEGED0018 | Dextran Sulfate Salt Detection kit |
Host Cell DNA Detection kits (HCD) | |
NS-D050T/NS-D100T | NS0 Host Cell DNA (NS0 HCD) Residue Detection kit |
EC-D050T/EC-D100T | E.coli Host Cell DNA (E.coli HCD) Residue Detection kit |
VE-D050T/VE-D100T | Vero Host Cell DNA (Vero HCD) Residue Detection kit |
HK-D050T/HK-D100T | HEK293 Host Cell DNA (HEK293 HCD) Residue Detection kit |
CH-D050T/CH-D100T | CHO Host Cell DNA (CHO HCD) Residue Detection Kit |
PP-D050T/PP-D100T | Pichia Pastoris Host Cell DNA (PP HCD) Residue Detection Kit |
CG-DP050/CG-DP100 | Magnetic Residual DNA Sample Preparation Kit |
Residual Total RNA Detection Kit | |
EC-R100T | E.coli Residual Total RNA Detection Kit (qRT-PCR) |
Host Cell Protein Antibodies | |
CH-K0018-3-Ab | CHO Host Cell Protein G3 Antibody |
CH-K0018-2-Ab | CHO Host Cell Protein G2 Antibody |
EH-E0020-3-Ab | E.coli Host Cell Protein G3 Antibody |
PH-E0021-2-Ab | Pichia Yeast Host Cell Protein G2 Antibody |
HH-H0019-2-Ab | HEK293 Host Cell Protein G2 Antibody |
Buffer Products | |
CG-H0100 | HCP ELISA buffer |
CG-P0200 | Protein L ELISA buffer |
