1. Introduction to the Background

 

Biopharmaceuticals began to emerge in the 1970s with the advent of DNA recombinant technology and monoclonal antibody technology. The launch of the first recombinant insulin in 1982 marked the official beginning of the biopharmaceutical era. After more than 40 years of development, biopharmaceuticals now encompass a wide range of biological macromolecules and cell-based therapies, including antibody drugs, vaccines, recombinant protein drugs, cell therapies, and gene therapies.

 

The biopharmaceutical market is experiencing rapid growth. Taking the Chinese market as an example, the domestic biopharmaceutical market size increased from 45.2 billion US dollars in 2019 to 66.5 billion US dollars in 2023, with a compound annual growth rate (CAGR) of 10.13%. It is projected that by 2030, the Chinese biopharmaceutical market size will reach 162.8 billion US dollars, with an estimated CAGR of 13.64% from 2023 to 2030. Globally, the biopharmaceutical market had a CAGR of 9.0% from 2018 to 2023, reaching 402.1 billion US dollars in 2023. It is anticipated that the global biopharmaceutical market will have a CAGR of 7.5% from 2023 to 2030, with a projected market size of 665.1 billion US dollars by 2030.

 

Compared with traditional drugs, the advantages of biopharmaceuticals are primarily reflected in the following aspects:

1) Highly Specific Mechanism of Action: Biopharmaceuticals can bind to their targets with a high degree of specificity, akin to a key fitting into a lock. This enables them to exert therapeutic effects while minimizing damage to normal cells.

2) Significant Therapeutic Efficacy: Biopharmaceuticals are particularly effective in treating complex and chronic diseases, as well as conditions where traditional drugs have limited efficacy. They offer new hope and more effective therapeutic options for patients.

3) Lower Immunogenicity: When properly designed and manufactured, biopharmaceuticals exhibit lower immunogenicity compared to traditional chemical drugs, which may cause widespread side effects. This reduces the potential for interference with normal physiological functions in the human body.

 

Biopharmaceuticals have demonstrated substantial potential in therapeutic applications. However, their production processes are significantly more complex than those of small-molecule chemical drugs synthesized through chemical reactions. The manufacturing of biopharmaceuticals involves the engineering and cultivation of tool cells or bacteria. Minute variations in conditions such as temperature and nutrition during cell culture, as well as subtle differences in product processing, purification, storage, and packaging, can all have a pronounced impact on the quality, purity, biological characteristics, and clinical efficacy of the final product.

 

Owing to the marked differences in production processes, quality control of biopharmaceuticals faces a number of unique challenges, especially in the area of residual detection. For traditional chemically synthesized drugs, residual detection primarily focuses on reaction by-products and impurities. In contrast, biopharmaceutical production involves a multitude of complex factors, including host cell components, reagents used in production, and potential contaminants. As a result, the scope and complexity of residual detection are correspondingly increased. In addition to microbial contamination and endotoxins, biopharmaceuticals also require detection of key residuals such as host DNA, host proteins, antibiotics, metabolic waste products, and enzymes.

 

2. Residual Host Cell Detection

 

Biopharmaceuticals are often produced using engineered cell lines. Residual host cell components introduced during the production process are critical quality control parameters. Depending on the specific components being detected, the primary categories include host cell proteins (HCPs) and host cell DNA (HCD).

 

Residual Host Cell Protein (HCP) Detection:

- The Chinese Pharmacopoeia (2020 Edition) stipulates that for CHO cells, the residual HCP level should be less than 0.05% (equivalent to less than 500 ppm); for E. coli, the residual HCP level should be less than 0.01%.

- The United States Pharmacopeia (USP) <1132> chapter specifies that a highly sensitive method should be used to detect HCPs in pharmaceutical products, with the content being below the limit of detection (typically less than 100 ppm, i.e., the HCP content in 1 mg of total protein should be less than 100 ng, which is equivalent to less than 0.01%).

- The European Pharmacopoeia (EP) 2.6.34 mandates that the HCP content in biological products should be less than 0.1%.

- The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines indicate that sensitive and validated methods in accordance with ICH criteria should be employed to monitor residual HCPs, with the residual levels typically required to be less than 100 ppm.

 

Residual Host Cell DNA (HCD) Detection: Typically, the residual HCD level is required to be below a specific threshold per dosage unit, such as less than 10 ng per dose, to prevent potential hazards associated with exogenous DNA to the human body.

 

Abbkine provides complete and reliable HCP & HCD detection kits for commonly used tool cells, featuring high sensitivity and good stability, to facilitate quality control in the production process.

 

CatalogNumber	Product Name	Specification	Features
abs590005	CHO HCP ELISA Kit	96T	Detection of CHO K1 HCP with high-coverage antibodies, strong specificity, and high sensitivity
abs590016	E. coli Host Cell Protein (HCP) Residual Detection Kit	96T	Detection of E. coli BL21 HCP with high-coverage antibodies, strong specificity, and high sensitivity
abs60544	Host Cell Residual DNA (Magnetic Bead Method) Sample Pre-treatment Kit	100T	Broad applicability (extraction of multiple HCDs), convenient and rapid (supporting manual and automated processes, completed in 1 hour)
abs60543	Plasmid DNA Residual (qPCR) Detection Kit	100T	Compatible with Host Cell Residual DNA Sample Pre-treatment Kit；   High sensitivity: Linear range 300 pg/µL to 0.03 pg/µL；   High precision: Low concentration repeatability CV < 20%; Medium to high concentration repeatability CV < 15% ；   Linear standard: In accordance with national standards, comprehensive indicators, high standards, and stable and reliable results；    Rapid and efficient: Quick detection with results available within one hour
abs60545	HEK293 Residual DNA (qPCR) Detection Kit	100T
abs60546	293T Cell Residual DNA (qPCR) Detection Kit	100T
abs60547	Human Residual DNA (qPCR) Detection Kit	100T
abs60548	Hela Cell Residual DNA (qPCR) Detection Kit	100T
abs60549	E1A Residual DNA (qPCR) Detection Kit	100T
abs60550	E1A & SV40LTA Residual DNA (qPCR) Detection Kit	100T
abs60551	E. coli Residual DNA (qPCR) Detection Kit	100T
abs60552	E. coli Total RNA Residual (qPCR) Detection Kit	100T
abs60553	CHO Residual DNA (qPCR) Detection Kit	100T
abs60554	Vero Residual DNA (qPCR) Detection Kit	100T
abs60555	Pichia pastoris Residual DNA (qPCR) Detection Kit	100T
abs60556	PG13 Residual DNA (qPCR) Detection Kit	100T
abs60557	HEK293 Residual DNA Fragment (qPCR) Detection Kit	100T
abs60558	293T Cell Residual DNA Fragment (qPCR) Detection Kit	100T
abs60559	Human Residual DNA Fragment (qPCR) Detection Kit	100T
abs60560	Hela Cell Residual DNA Fragment (qPCR) Detection Kit	100T
abs60561	Vero Residual DNA Fragment (qPCR) Detection Kit	100T

 

3. Residual Process Additives Detection

 

The exogenous additives introduced during the production process of biopharmaceuticals must be evaluated and their residual levels detected to avoid adverse effects on humans due to high concentrations of these additives.

 

Culture Additives—BSA: Bovine serum albumin (BSA) is a common additive in cell culture processes. If used in the production process, the BSA content in the biopharmaceutical product must be detected. For instance, the Pharmacopoeia (2020 Edition) stipulates that the residual BSA level in the final vaccine product should not exceed 50 ng/mL or 50 ng per dose.

 

Culture Additives—Antibiotics: Antibiotics are commonly employed in cell culture to prevent contamination or for cell selection processes. When used in the production of biopharmaceuticals, efforts should be made to remove antibiotics from the final product to the greatest extent possible. The Pharmacopoeia (2020 Edition) of China stipulates that if antibiotics are added during the production process of biopharmaceuticals, the residual levels must be tested. The cumulative residual amount of all antibiotics in the final product should not exceed 50 ng per dose.

 

Production-Related Residuals—Protein A: Protein A is a key reagent in the affinity purification process of antibodies and is commonly used in the affinity chromatography purification step of monoclonal antibody (mAb) production. Due to factors such as material aging, Protein A immobilized on the purification matrix may detach and enter the purified antibody, resulting in residual contamination. Pharmacopoeias also specify the residual limits for Protein A. For instance, in the 2020 Edition of the Pharmacopoeia, Part III, under the section for “Injection of Nimotuzumab” (3.1.3.3), it is stipulated that the residual Protein A level should not exceed 0.001% of the total protein content, as determined by the enzyme-linked immunosorbent assay (General Rule 3429).

 

Production-Related Residuals—RNase Inhibitor: During the production process of mRNA vaccine drugs, RNase Inhibitor effectively suppresses the RNase activity that may be present in the reaction system, thereby protecting mRNA from degradation. According to regulations, various residual tests must be conducted on the bulk solution of mRNA vaccines. As a protein product, RNase Inhibitor falls within the scope of protein residual detection. Therefore, its residual level must be quantitatively determined.

 

Production-Related Residuals—RNA Polymerase: During the preparation of mRNA, T7 RNA polymerase is one of the key raw materials and serves as the core enzyme in the development of mRNA vaccines and therapeutics. However, for the final drug product, T7 RNA polymerase is considered an impurity. T7 RNA polymerase, when encapsulated with mRNA into lipid nanoparticles (LNPs) and released into cells, can act as a foreign antigen. It can bind to complementary antibodies, induce pro-inflammatory cytokines as part of the adaptive immune response, and lead to inflammation, which in turn may trigger the degradation of mRNA. Therefore, more robust assessment methods are required to detect its residual levels, ensuring the quality and safety of the final product.

 

Production-Related Residuals—Collagenase: Collagenase is utilized for dissociating a variety of tissues into single cells in vitro and is applicable for the isolation of single cells from organs and tissues for diverse research purposes. Regulatory guidelines pertinent to biopharmaceutical production have underscored the necessity for testing materials associated with cell culture. Consequently, the detection of residual collagenase levels has garnered increasing attention.

 

Production-Related Residuals—Antibodies: During the production of cell therapy drugs such as CAR-T, antibody-related impurities may be introduced at various stages, including cell sorting and activation, viral transduction, and purification. Therefore, the detection of residual antibody levels also requires attention.

 

To address these requirements, Abbkine offers dedicated detection kits for the aforementioned indicators, thereby meeting the needs of customers.

 

CatalogNumber	Product Name	Specification	Features
abs590008	BSA ELISA Kit	96T	Strong specificity, no cross-reactivity with other species; high sensitivity, detection limit < 0.5 ng/mL; high accuracy, good reproducibility
abs590010	Kanamycin Residual Detection Kit	96T	High sensitivity, detection limit < 0.05 ng/mL; high accuracy, good reproducibility; two-step competitive method, results in 1 hour, convenient and rapid
abs590011	Gentamicin Residual Detection Kit	96T
abs590009	Protein A Residue Detection Kit	96T	High sensitivity, not affected by high concentrations of IgG
abs520031	RNase Inhibitor ELISA Kit	96T	Sandwich ELISA, high sensitivity, good reproducibility
abs590012	Collagenase Type I Tesidue Detection Kit	96T	Sandwich ELISA, high sensitivity, good reproducibility
abs590015	T7 RNA Polymerase Residual Detection Kit (ELISA Method)	96T	Sandwich ELISA, high sensitivity, good reproducibility
abs560002	Human Total IgG Kit (Assay Pro)	100-10000T	Time-resolved fluorescence (TRF) technology, homogeneous no-wash detection, competitive method, rapid, sensitive
abs560011	Human IgG Sandwich Assay Kit	100-10000T	Time-resolved fluorescence (TRF) technology, homogeneous no-wash detection, sandwich method, rapid, sensitive
abs560041	Human IgG Kit (HICA)	200-5000T	Homogeneous chemiluminescence technology, homogeneous no-wash detection, chemiluminescence, ultra-wide linear range, high sensitivity, rapid
abs551021	Human IgG ELISA Kit	96T	Traditional sandwich ELISA, a classic detection method

 

4. Residual Contaminants Detection

 

During the production and preparation of biopharmaceuticals, it is imperative to rigorously identify and eliminate all potential contaminants to prevent adverse effects on human health. Common contaminants in the production process include mycoplasma contamination that may occur during the cultivation of tool cells and endotoxin contamination that can arise throughout the entire production process. To address these concerns, the Pharmacopoeia explicitly mandates that biopharmaceuticals must undergo mycoplasma testing, with a detection sensitivity of no less than 10 colony-forming units (CFU) per milliliter. Similarly, for endotoxins, the Pharmacopoeia provides detailed regulations. For recombinant cytokine biopharmaceuticals such as interleukins, interferons, and tumor necrosis factors, the endotoxin limit is typically between 1 and 5 endotoxin units (EU) per dose. In contrast, different vaccines have varying endotoxin limits. For instance, the recombinant hepatitis B vaccine (Saccharomyces cerevisiae) requires that the endotoxin content per dose be strictly controlled to less than 5 EU.

 

CatalogNumber	Product Name	Specification	Features
abs9925	qPCR Mycoplasma Test Kit	50T	High Detection Rate: The detection rate for 10 standard mycoplasma strains mentioned in various pharmacopoeias, including oral mycoplasma, porcine mycoplasma, and pneumonia mycoplasma, is 100% at a concentration of 10 CFU/mL. No false positives, good reproducibility, with a CV value below 5%.
abs90283	MYCO-L™ Luminescent Mycoplasma Detection Assay Kit	10T/25T/50T	Broad Detection Range: The detection range is extensive, with sensitivity comparable to that of qPCR methods. Detection is rapid, with results available within 30 minutes.
abs500001	Recombinant C Factor Endotoxin Detection Kit	48T/96T	Recombinant C Factor as an alternative to traditional Limulus Amebocyte Lysate (LAL), with a detection limit as low as 0.005 EU/mL, in accordance with pharmacopoeial methods.

 

5.Impurities Detection

 

Owing to the unique nature of their production processes, biopharmaceuticals may generate specific by-products that necessitate particular detection protocols throughout the entire manufacturing process. For instance, during the production of mRNA vaccines, single-stranded mRNA may aggregate to form double-stranded RNA (dsRNA) impurities. Similarly, following the induction of CAR-T cells, these cells secrete inflammatory cytokines, which must be removed during the production process, with the efficiency of removal also being subject to detection.

 

CatalogNumber	Product Name	Specification	Features
abs590014	dsRNA ELISA Kit	96T	Specific detection of dsRNA longer than 60 bp, independent of sequence; compatible with detection of dsRNA containing various modified bases; high sensitivity and good reproducibility.
abs560022	Human IL-2 Kit (HICA)	500-10000T	Homogeneous chemiluminescence technology, no-wash homogeneous detection, chemiluminescence, ultra-wide linear range, high sensitivity, rapid.
abs510028	Human IL-7 ELISA Kit	96T	Traditional Sandwich ELISA, a Classic Detection Method
abs510018	Human IL-15 ELISA Kit	96T	Traditional Sandwich ELISA, a Classic Detection Method
abs551810	Human IL-21 High Sensitivity ELISA Kit	96T	High-sensitivity ELISA with TSA (Tyramide Signal Amplification) for Lower Detection Limits
abs50186	Human 12 Cytokines Multiplex Assay Kit (CBA)	48T/96T	CBA (Cytometric Bead Array) Flow Cytometry Multiplex Assay: Detection of 12 Parameters from a Single Sample

 

Absin provides antibodies, proteins, ELISA kits, cell culture, detection kits, and other research reagents. If you have any product needs, please contact us.

Absin Bioscience Inc. Email: worldwide@absin.net

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