Biopharmaceuticals and biotherapeutics are medicines produced from living cells. But cells produce other proteins (called host cell proteins, or HCPs) that can contaminate the final therapeutic product and cause a number of problems.
Download this infographic to learn about:
- What HCPs are and the problems they pose
- Established approaches for HCP analysis
- New and emerging HCP analysis approaches
Type of Drug Dose Frequency of administration Disease
However, these cells also make other proteins alongside the desired
therapeutic product. These are known as host cell proteins (HCPs).
HCPs are a major source of contamination as they can persist through the
manufacturing process and contaminate the final therapeutic product.
This can pose a number of problems:
A biopharmaceutical must meet the acceptable levels of HCPs as
required by agencies such as the US Food and Drug Agency (FDA) and
European Medicines Agency (EMA).
The acceptable level is usually evaluated on a case-by-case basis, and
depends on factors such as:
HCP analysis is not usually achieved using a single tool, but several
orthogonal methods.
There are several approaches commonly used in the detection,
characterization and identification of HCPs.
The gold-standard approach and most common method, semiquantitative and widely accepted by regulatory agencies.
Triggering immune responses in patients
Affecting the drug’s efficacy
Reducing drug stability and quality
Biopharmaceuticals and biotherapeutics (also known as
biologics) are medicines produced from living cells.
Antibodies
Vaccines
Cell and gene therapies
Approaches & Advances
include:
Host Cell Protein
Analysis:
HCP analysis
severity
Approaches for
An orthogonal approach is a different technique that
measures the same property using a different method.
This helps to confirm the original result and eliminate
false positives.
Examples
HCP analysis
Established
approaches for
ELISA enzyme-linked
Method:
Antibodies are immobilized on a plate and the samples
added. HCPs bind specifically to antibodies. Detection
antibodies then bind, forming a “sandwich” that can be
used to quantify the amount of protein present.
SUBSTRATE
SECONDARY
ANTIBODY
EXAMPLE OF SANDWICH ELISA
DETECTION
ANTIBODY
CAPTURE
ANTIBODY
HCP
Pros &Cons:
Relatively simple, sensitive, well-validated and selective.
High throughput, analyzing many samples in a short period.
Can only detect HCPs that have specific antibodies available
and may miss HCPs not recognized by the antibody mix.
Only provides information on total HCPs.
Separates proteins based on isoelectric point and molecular weight
across a polyacrylamide gel to allow for identification of individual HCPs.
Recommended to characterize HCP antigens and anti-HCP antibodies.
2D gel electrophersis
Method:
Biochemically characterizes HCPs in a sample by
separating according to isoelectric point (pI) in one
dimension and molecular weight (MW) in the other.
pl
MW
HIGH MW + LOW pl
LOW MW + LOW pl
HIGH MW + HIGH pl
LOW MW + HIGH pl
Pros &Cons:
High-resolution separation allows for investigation of HCP
population all at once.
Allows characterization of intact proteins.
Labor-intensive with low throughput and low reproducibility.
Provides limited quantitative data and is highly techniquedependent – needs to be combined with other techniques.
Mass spectrometry-based approaches are the main emerging techniques
that are currently rising in popularity.
A newer method that is increasingly being used as an orthogonal
approach to ELISA. It has become a key HCP analysis tool for the
identification and quantification of individual proteins.
HCP analysis
Emerging
technologies for
Molecules in a sample are ionized and broken down
into smaller peptides. They are typically separated by
liquid chromatography (LC) and then analyzed by MS,
by their mass-to-charge ratio.
spectrometry (LC-MS)
Liquid chromatography-mass
Method:
BIOLOGIC WITH HCP
IMPURITIES
PEPTIDE
FRAGMENTS
LC SEPARATION
OF PEPTIDES MS ANALYSIS
Pros &Cons:
More comprehensive detection that distinguishes between
various types of HCP. Can also detect low-abundance
proteins that ELISA may miss.
Less high-throughput, more complex and more expensive,
requiring technical expertise and equipment.
Limited dynamic range, difficult to detect low-abundance
proteins.
Future perspectives
The future of HCP analysis may bring improved assays and HCP clearance
as well as changes in expression systems to reduce HCP expression. HCP
analysis is essential for the safety and efficacy of biopharmaceuticals,
helping to detect impurities that could impact patient health. Both
existing methods such as ELISA and emerging approaches like LC-MS can
help biopharma companies to profile HCPs. As these tools continue to
evolve and develop, this can help us to produce safer and more effective
therapies for patients.
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