OUR METHODS Service Workflow Platforms Why Choose US? FAQs Resources
Bispecific antibodies (BsAbs) exhibit enhanced therapeutic effects over traditional monoclonal
antibodies. However, achieving a high-quality bsAb product requires not only expert design and
engineering but also robust and reliable purification processes to meet clinical standards. Due to
the structural complexity and heterogeneity of bsAbs, purification presents unique challenges. The
production of bsAbs often results in unwanted forms, including mismatched chains, aggregates, and
non-functional variants, making separation and purification difficult. Advanced strategies are
required to isolate correctly assembled bsAbs from byproducts that exhibit similar physicochemical
properties.
At Creative Biolabs, we address these challenges with specialized purification techniques optimized
for each client's configuration of symmetric, asymmetric, or fragment-based bsAbs. Our services
include affinity chromatography-based capture, and charge, size, hydrophobicity and mixed-mode-based
separation methods to ensure high purity and yield. With extensive experience in handling complex
biologics, our team ensures that purified bsAbs are compatible with downstream processes such as
formulation and sterile filtration, facilitating seamless transitions to clinical manufacturing.
OUR Methods
Creative Biolabs employs a customized combination of purification methods, including capture steps
and polishing procedures, to optimize the success of each project. Regardless of the bsAb format,
our optimized workflow guarantees high purity and yield, meeting the specific requirements of each
product. With extensive expertise in handling various bsAb configurations, we ensure efficient
purification that supports seamless downstream applications and accelerates project timelines.
Type of Affinity Chromatography
|
Principle
|
Protein A
|
Protein A specifically binds to the Fc region of IgG antibodies, facilitating separation
of
IgG from non-antibody components. Binding is typically stable at neutral pH, while
elution
is achieved by lowering pH.
|
Protein G
|
Protein G binds specifically to the Fc regions of IgG subclasses and certain polyclonal
antibodies. It offers higher binding affinity for certain IgG subclasses than Protein A.
|
Protein L
|
Protein L binds to the kappa light chains of antibodies, making it suitable for
purifying
Fab fragments, scFvs, and other non-Fc antibodies.
|
HRPO (Horseradish Peroxidase)
|
HRP is conjugated to antibodies for detection purposes. It reacts with substrates like
TMB
or DAB, creating a colorimetric or chemiluminescent signal, which can be used to track
bispecific antibodies during purification or in bioassays.
|
IMAC (Immobilized Metal Affinity Chromatography)
|
Metal ions (e.g., Ni²⁺, Co²⁺) immobilized on the resin bind to histidine residues on the
target protein or antibody fragment. Elution is achieved with imidazole or acidic
buffers.
|
Charge-based separation (Ion-Exchange Chromatography - IEX)
|
Charged molecules bind to oppositely charged ligands on the stationary phase. Elution is
achieved by changing the pH or ionic strength of the mobile phase.
|
Size-based separation (Size-Exclusion Chromatography - SEC)
|
Molecules are separated based on their size as they pass through a porous matrix. Small
molecules enter the pores, while large molecules are excluded and elute faster.
|
Hydrophobicity-based separation (Hydrophobic Interaction Chromatography - HIC)
|
Hydrophobic proteins bind to a hydrophobic resin in high salt concentrations. Elution
occurs
by decreasing the salt concentration, reducing hydrophobic interactions.
|
Mixed-mode-based purification (Mixed-Mode Chromatography - MMC)
|
Mixed-mode ligands interact with target molecules through hydrophobic, ionic, and
hydrogen
bonding simultaneously. Elution is achieved through changes in salt concentration or pH.
|
SERVICE WORKFLOW
Project Consultation and Design
- Collaborate with clients to understand the specific bispecific antibody (BsAb)
format and project
requirements.
- Design a tailored purification strategy based on antibody structure, binding
properties, and production
scale.
- Establish timelines and quality control checkpoints for the purification process.
Sample Preparation
- Receive BsAb samples from clients or coordinate production within Creative Biolabs'
facilities.
- Perform initial quality checks (e.g., concentration, aggregation analysis) to ensure
the sample is ready
for purification.
Capture Step – Primary Purification
- Use affinity chromatography (e.g., Protein A, Protein G, or IMAC) to isolate the
target bispecific
antibody.
- Remove major impurities, including host cell proteins, DNA, and process-related
contaminants.
- Collect eluate fractions for further processing.
Polishing Step – Secondary Purification
- Apply advanced purification techniques such as Ion-Exchange Chromatography (IEX),
Size-Exclusion
Chromatography (SEC), or Hydrophobic Interaction Chromatography (HIC).
- Refine the product to remove aggregates, isoforms, and residual impurities.
Quality Control and Analysis
- Conduct rigorous analytical testing, including SDS-PAGE, HPLC, mass spectrometry,
and bioassays, to verify
purity and functionality.
- Provide clients with a detailed certificate of analysis (COA) for each purified
batch.
Packaging and Delivery
- Package purified antibodies according to regulatory and client-specific guidelines.
- Coordinate secure and temperature-controlled shipping to ensure product integrity
upon delivery.
PLATFORMS
At Creative Biolabs, we utilize advanced instrumentation to offer precise and scalable
bispecific antibody purification services. Our platforms, including but not limited to the
ÄKTA avant™ chromatography system, Q Exactive™ Plus mass spectrometry, and Agilent Infinity
III HPLC, are tailored to address the complexities of various bsAb formats, ensuring high
purity and optimal yields. Our comprehensive platform validation and quality control
processes guarantee consistent results and meet the stringent requirements of regulatory
authorities.
Each phase of the purification process, from automated chromatography to high-resolution mass
spectrometry and buffer exchange, is powered by state-of-the-art equipment. These tools
enable real-time monitoring, detailed characterization, and seamless scalability, ensuring
reliable performance throughout research and clinical production stages. Additionally, our
integrated data analysis capabilities and dedicated technical support ensure optimal process
parameters and reproducible results for every project.
WHY CHOOSE US?
At Creative Biolabs, we offer a unique blend of expertise, innovation, and customization for
bispecific antibody purification services, making us an experienced partner for your bsAb
development needs.
Comprehensive Expertise Across bsAb Formats
Creative Biolabs has extensive experience in handling diverse bispecific antibody
formats. Each format presents unique structural challenges, but our tailored
purification strategies ensure optimal recovery, purity, and functionality,
regardless of the complexity of the molecule.
Customized Two-Step Purification Process
We implement a robust two-step purification process for every project. The first
capture step isolates the target bispecific antibody, which is followed by polishing
steps to remove aggregates, byproducts, and residual impurities, ensuring
high-purity antibodies suitable for clinical applications.
Advanced Purification Technologies and Analytical Tools
Our state-of-the-art purification platforms employ advanced chromatography
techniques, including mixed-mode purification for challenging protein mixtures.
Complemented by sophisticated analytical methods such as HPLC, we guarantee the
integrity and consistency of the final product.
Optimized Yield Without Compromising Quality
We understand the importance of both yield and purity in antibody production. Our
processes are optimized to maximize recovery without compromising on quality,
ensuring that clients receive high-performing bispecific antibodies that meet
regulatory and application-specific standards.
Rigorous Quality Control and Documentation
Every purified batch undergoes thorough quality control, with detailed testing to
confirm purity, functionality, and stability. We provide a comprehensive certificate
of analysis (COA) with every delivery, ensuring transparency and compliance with
industry standards.
Rapid Turnaround with Flexible Scale-Up Options
Our streamlined purification workflows and dedicated project management ensure quick
turnaround times without compromising quality. We offer flexible scale-up options
from small-scale process development to large-scale production, supporting clients
through different stages of their development pipeline.
FREQUENTLY ASKED QUESTIONS
What information or materials do I need to
provide to start the CAR design process?
To initiate the CAR design and construction process, we typically require a few key pieces
of information. First, we need details about the specific target antigen or antigens you
want to address. If you have any data on the antigen’s expression profile or binding
characteristics, that would be extremely helpful. Additionally, sharing any preferences
regarding the structure of the CAR (e.g., co-stimulatory domains, signaling domains, or
transmembrane regions) allows us to customize the design more effectively. If you have
existing sequences for the antigen-binding domain (such as an scFv) or previous constructs
you’ve worked on, providing those sequences can help accelerate the process. If you’re
starting from scratch, don’t worry—we can also assist with identifying the appropriate
components for your CAR based on your therapeutic or research goals.
What deliverables can I expect from your CAR
design and construction service?
Our CAR design and construction service provides a comprehensive set of deliverables to
ensure you have everything you need to move forward with your project. You will receive a
fully optimized CAR construct that includes the gene sequence of the entire CAR, designed
according to your specifications. We also offer gene sequence optimization to improve
expression and reduce potential immunogenicity. Depending on your needs, we can provide the
CAR in different vector formats, such as plasmid, lentiviral, or adenoviral vectors, which
are fully validated for functionality. Additionally, you will receive detailed reports on
sequence verification (e.g., Sanger or next-generation sequencing), expression validation
(using methods like flow cytometry or Western blot), and functional testing results, such as
cytotoxicity assays. These deliverables give you a fully functional and validated CAR, ready
for further in vitro or in vivo studies.
Can I select specific services or customize
the package based on my needs?
Yes, our services are fully customizable to fit the unique requirements of your project.
Whether you need only a specific aspect of the CAR-T development process—such as the design
of the CAR structure, vector construction, or validation testing—we can provide standalone
services. For example, if you have already designed your own CAR but need assistance with
cloning and vector construction, we can handle that part of the process. Alternatively, if
you need a complete solution from design to functional validation, we can offer an
end-to-end service. You are not locked into a one-size-fits-all package. We encourage
clients to discuss their specific goals and challenges with our team so we can create a
custom service plan that suits their budget and timelines.
How will I be involved in the design and
development process?
Our approach to CAR design and construction is highly collaborative, ensuring that you are
actively involved at every critical step of the process. From the initial consultation, we
work closely with you to understand your project’s objectives, therapeutic targets, and any
preferences you have for the CAR’s structure. Throughout the design phase, we will provide
regular updates and involve you in key decisions, such as selecting co-stimulatory domains,
antigen-binding regions, and vector options. As the project progresses, you will receive
milestone reports, and we will schedule review meetings to discuss any necessary
adjustments. This level of engagement ensures that the final CAR construct fully aligns with
your expectations and research goals. Your feedback is valuable at every stage, and our team
is always available to answer questions and make modifications as needed.
Are there options to scale up the production
of CAR constructs after initial design and
validation?
Yes, we offer scalable solutions that can support your project as it moves from research and
development into preclinical or clinical phases. Once your CAR design has been finalized and
validated, we can assist with scaling up production based on your specific needs. For
example, if you require large-scale plasmid production for further in vitro testing
or therapeutic applications, we offer high-quality plasmid preparation in various purity
grades. Additionally, we provide viral packaging services, such as lentiviral and adenoviral
production, which are suitable for both small-scale studies and larger-scale applications.
Our team can help you transition smoothly to higher quantities of CAR constructs for
preclinical studies or clinical-grade production. Furthermore, we ensure that all
large-scale products undergo the same rigorous quality control and validation processes to
maintain consistency and reliability as your project scales.
RELATED RESOURCES
"We worked with Creative Biolabs to purify an IgG-like bispecific antibody from our CHO cell culture supernatant. The sample had significant aggregation issues, but their team applied SEC as a polishing step, and the final product was over 98% pure. What stood out was their ability to recover a high yield despite initial challenges. The detailed Certificate of Analysis (COA) they provided gave us complete confidence in the product’s quality."
— Dr. Sarah Mitchell – Principal Scientist