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Chimeric Antigen Receptor (CAR) Design and Construction Services

Discover the power of CAR technology with tailored design and construction services to enhance precision, safety, and therapeutic impact. From innovative constructs to advanced generations, Creative Biolabs is committed to driving your research forward and accelerating your journey to clinical breakthroughs.

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Chimeric antigen receptor (CAR) T cell therapy represents a groundbreaking advancement in the fight against cancer and other diseases. However, each step, from selecting appropriate antigen-binding domains to optimizing CAR structural components, is crucial for developing CARs with high specificity and effectiveness. Understanding these complexities, we are committed to providing comprehensive CAR design and construction services tailored to meet the unique requirements of each project. Whether it's conventional or novel designs, our experienced team employs innovative thinking to ensure each CAR we help construction maximizes efficacy and safety.

Utilizing advanced bioinformatics tools and molecular biology techniques, we guide clients through the complex CAR development process from initial conceptualization to final validation. Our services are designed to accelerate research and development timelines, facilitating swift transitions from the lab to clinical applications. Whether your focus is on hematologic malignancies, solid tumors, or exploring CAR applications in autoimmune diseases, our goal is to support your work by providing cutting-edge solutions and expert guidance. Partner with us to access rich knowledge and resources dedicated to advancing the frontier of cellular immunotherapy.

Fig. 1 Chimeric antigen receptor design and construction services. (Creative Biolabs Authorized)

Chimeric Antigen Receptor Design

Fig. 2 Basic structure of a chimeric antigen receptor. (Creative Biolabs Original)
  • CAR components design
  • DIY your own design

CAR Construction & Validation

Fig. 3 Chimeric antigen receptor design. (Creative Biolabs Original)
  • Gene sequence design and optimization
  • Vector selection

Verification and Validation

Fig. 4 CAR vector construction. (Creative Biolabs Original)
  • Sequence verification
  • Expression validation
  • Stability and functional testing

Chimeric Antigen Receptor Design

At the forefront of CAR-T therapy development, our CAR Design services aim to create highly effective and targeted therapies for various cancers. By leveraging advanced design strategies, we ensure that each component of the CAR is optimized for peak performance. Our approach encompasses both established and innovative CAR formats tailored to meet specific therapeutic needs. Additionally, we prioritize humanization and low immunogenicity to enhance the safety and effectiveness of our CAR constructs.

CAR components design

Fig. 5 CAR cell verification and validation. (Creative Biolabs Original) Antigen-binding region (scFv)
We design scFvs with high affinity and specificity to the target antigen. Our team further optimizes scFv sequences through computational modeling, enhancing stability and reducing immunogenicity. Learn more about our ScFv Generation Services.
Fig. 6 Antigen-binding region (scFv) of a chimeric antigen receptor. (Creative Biolabs Original)Hinge and trans-membrane domain
We adjust the hinge region based on antigen spatial distribution, ensuring optimal flexibility and activity during antigen binding. We offer a range of transmembrane domain options with enhanced CAR surface expression and function.
Fig. 7 Hinge and trans-membrane domain of a chimeric antigen receptor. (Creative Biolabs Original)Co-stimulatory and signaling domain
By incorporating different generations of co-stimulatory domains, we can fine-tune the strength, duration, and safety of T cell activation. This flexibility allows us to customize CAR designs for a variety of therapeutic applications.
Different generations of CAR designs
Fig. 8 Structural diagram of the 1st-generation chimeric antigen receptor. (Creative Biolabs Original)
1st Generation CAR
The original CAR-T or the first-generation CAR consists of the intracellular domain from the CD3 ζ- chain and the primary transmitter of signals from endogenous TCRs Although the anti-tumor activity was limited due to insufficient activation, persistence and homing to the cancer tissue, some significant effects indeed existed in patients with B-cell lymphoma treated with α-CD20-CD3 ζ CAR-modified T cells and some neuroblastoma treated with ScFv-CD3 ζ CAR-Ts. This is the most common form of CARs to fuse single-chain variable fragments (scFv) derived from monoclonal antibodies to CD3 ζ transmembrane and endo domain.
Fig. 9 Structural diagram of the 2nd-generation chimeric antigen receptor. (Creative Biolabs Original)
2nd Generation
The 2nd-generation CARs were designed to combine the intracellular signaling domains from various costimulatory protein receptors (e.g., CD28, 41BB, ICOS) incorporated in the cytoplasmic tail of the CAR to enhance the signaling. For instance, the CD19-targeted CARs incorporated with CD28 or 4-1BB signaling domains manifested remarkable complete remission rates in patients with refractory B-cell malignancies. Subsequently, the CD28-based CARs showed a brisk proliferative response and boost effector functions. Meanwhile, the 4-1BB-based CARs manifested a more progressive T cell accumulation.
Fig. 10 Structural diagram of the 3rd-generation chimeric antigen receptor. (Creative Biolabs Original)
3rd Generation
As the expectation of more antitumor efficacy, the 3rd-generation of CARs combined multiple signaling domains (e.g., CD3 ζ-CD28-41BB) to acquire further enhanced activation signals, proliferation, production of cytokines and effective function. For instance, the α-CD19-CD3 ζ-4-1BB CAR-Ts for chronic lymphocyte leukemia showed complete remission to infiltrate and lyse cancer tissue. Even better, a fraction of CAR-Ts functioned as a memory phenotype for preventing tumor relapses. While the emerging uncontrollable activity accompanied with more antitumor efficacy caused life-threatening lysis activity as the most critical adverse effect.
Fig. 11 Structural diagram of the 4th-generation chimeric antigen receptor. (Creative Biolabs Original)
4th Generation
The previous CAR strategies are highly specific and useful in redirecting T cells targeting malicious cancer cells. However, the major limitation on solid tumors with a tremendous phenotypic heterogeneity and relapse due to antigen-negative cancer cells is the huge challenge to trigger a novel CAR strategy. The 4th-generation CAR is designed to shape the tumor environment by the inducible release of transgenic immune modifiers, such as IL-12, which augments T-cell activation, attracts and activates innate immune cells to eliminate antigen-negative cancer cells in the targeted lesion.
Fig. 12 Structural diagram of the 5th-generation chimeric antigen receptor. (Creative Biolabs Original)
5th Generation
The fifth generation of CARs, builds upon the second generation but includes a truncated cytoplasmic interleukin-2 receptor β-chain domain (IL-2Rβ) with a binding site for the signal transducer and activator of transcription-3 (STAT3). The antigen-specific activation of this receptor triggers three critical signaling pathways simultaneously: TCR signaling through the CD3ζ domains, co-stimulatory signaling via the CD28 domain, and cytokine signaling through the JAK–STAT3/5 pathway. This integrated approach provides the synergistic signals necessary for full T cell activation and proliferation, effectively enhancing the immune response.
Innovative CAR formats
Fig. 13 Structural diagram of Dual CAR. (Creative Biolabs Original)
Dual CAR
Dual CAR T cells improve the specificity and safety of CAR-T therapy by targeting two tumor-associated antigens simultaneously. They are designed to activate only when both antigens are present on the tumor, reducing off-tumor toxicity. These cells express two CARs: one binding to a specific tumor antigen and delivering the primary signal, and another binding to a different antigen and providing the co-stimulatory signal. For example, dual CAR-T cells co-expressing anti-ErbB2 and anti-MUC1 CARs use CD3ζ and CD28 signaling pathways, respectively, and effectively kill ErbB2+ tumor cells only in the presence of MUC1.
Fig. 14 Structural diagram of Conditional CAR. (Creative Biolabs Original)
Conditional CAR
Conditional CAR enhances the safety of CAR-T cell therapy by requiring an additional signal which can be a small molecule or an adaptor-specific receptor that binds to secondary antibodies directed at target cell antigens. It acts as a switch, activated by an exogenous molecule, allowing precise regulation of CAR-T cell activity without leading to cell death in its absence. For example, a conditional CAR targeting CD19 in a Nalm-6 xenograft rodent model demonstrated controlled activity, tissue-homing, cytokine release, and phenotype in a dose-titratable manner, potentially reducing CRS compared to conventional CAR-T cells.
Fig. 15 Structural diagram of TanCAR. (Creative Biolabs Original)
TanCAR
TanCARs are an advanced type of chimeric antigen receptor designed to enhance the efficacy of CAR-T cell therapy. Unlike traditional CARs, TanCARs feature two distinct antigen recognition domains on a single transgenic receptor. This allows for simultaneous targeting of tumor cells and elements within the tumor microenvironment, increasing T cell activation and function by enhancing avidity and broadening therapeutic reach. This dual targeting capability addresses issues like antigen-loss escape variants commonly seen in cancer cells, potentially leading to more effective and durable cancer treatments.
Fig. 16 Structural diagram of Physiological CAR. (Creative Biolabs Original)
Physiological CAR
Unlike traditional CARs, which often use scFv of murine origin and risk triggering immune responses, physiological CARs utilize receptor ligands such as HER3 and HER4. These CARs consist of an antigen receptor and a CD3ζ intracellular signaling domain, with or without a transmembrane and spacer region, engineered into immune cells to target ligands expressed on tumor cells. This approach enhances the ability of T cells to recognize and eliminate cancer cells, increasing the persistence and effectiveness of the therapy while reducing the risk of immune response and anaphylaxis associated with murine-based scFv CARs.
Fig. 17 Structural diagram of Universal CAR. (Creative Biolabs Original)
Universal CAR
Universal CARs represent a significant advancement in CAR-T cell therapy by offering flexibility and adaptability to target a wide range of malignancies. These CARs are engineered to recognize various tagged antigens, such as biotin-labeled molecules, FITC-labeled targets, and peptide nucleic acids, among others. This versatility allows for precise and personalized treatment options, enhancing T cell activation, controlling therapeutic effects, and reducing adverse reactions. Universal CARs improve the scope and efficacy of CAR-T therapies, making them more effective against diverse cancer types.
Fig. 18 Structural diagram of Marked CAR. (Creative Biolabs Original)
Marked CAR
Marked CARs are specially designed CARs that express both CARs and tumor epitopes that are recognized by existing monoclonal antibodies. This dual expression allows for a unique safety mechanism: if severe adverse effects occur, the specific monoclonal antibody can be administered to selectively clear the CAR-T cells, thereby avoiding additional off-tumor effects. This approach not only helps in mitigating side effects but also enhances the overall therapeutic potential by providing a controlled and targeted method to manage and eliminate CAR-T cells if necessary. In essence, Marked CAR combines efficacy with improved safety.

These designs focus on modifying the basic structural components of CARs to optimize their binding and recognition capabilities.
Categories Designs Features
Specific Binding Domain Modifications VHH-based CAR Construction Uses single-domain antibodies derived from camelids, offering smaller size and better tissue penetration.
Adnectin-Based CAR Construction Employs engineered fibronectin type III domains as targeting molecules, providing high stability and specificity.
Peptide-centric CAR Construction Utilizes synthetic peptides as targeting domains, allowing for highly customizable antigen recognition.
TCR-fused Antigen Binding Receptor (TCR-ABR) CAR Construction Combines TCR and CAR technologies to enhance recognition of intracellular antigens.
CD3ε-Based CAR Design & Construction Incorporates CD3ε signaling components to improve T cell activation and signal transduction.
Immune Receptor Integration NKG2D-based Multi-target CAR Construction Service Utilizes natural killer cell receptors to target stress-induced ligands on tumor cells.
KIR CAR Construction Incorporates killer cell immunoglobulin-like receptors for enhanced tumor recognition.
γδ CAR-T Cell Development Combines γδ T cell properties with CAR technology for broader antigen recognition.
High-Efficiency Genomic Integration Use mammalian-derived genome engineering enzymes to flawlessly introduce DNA of any size into precisely defined genomic locations.
CHAR Construction CHAR construction to improve the elimination of B cells producing donor-specific HLA antibodies
These designs incorporate various control mechanisms to regulate CAR-T cell activity and enhance safety profiles.
Categories Designs Features
Controllability Designs Smarter™ Self-destruct CAR Construction Includes a suicide switch mechanism for controlled elimination of CAR-T cells if needed.
Smarter™ Switch CAR-T Construction Features an on/off switch system for regulated CAR-T cell activity.
FR806-based Switchable CAR Construction Uses small molecule-dependent activation for controlled CAR-T cell function.
Smarter™ Orthogonal Dual-Switch CAR Construction Incorporates two independent control switches for enhanced safety and specificity.
Smarter™ Chemically Programmed Switch CAR Construction Enables chemical regulation of CAR-T cell activity through specific molecular triggers.
Logic Gate Control Logic-gated CAR Construction Uses Boolean logic gates to create complex activation requirements.
Modular CAR (modCAR) Construction Features interchangeable components for customizable CAR function.
These designs aim to improve the overall performance and effectiveness of CAR-T cell therapy.
Categories Designs Features
Functional Enhancement Smarter™ Armored CAR Construction Includes additional protective features to enhance CAR-T cell survival in hostile tumor environments.
Smarter™ 7 × 19 CAR Construction Optimized design incorporating multiple improvements for enhanced efficacy.
c-Jun Overexpressed CAR Construction Features increased c-Jun expression to improve T cell function and persistence.
IL-8R-modified CAR Construction Incorporates IL-8 receptor modifications for better tumor trafficking.
Chimeric Costimulatory Antigen Receptor-T Construction Includes enhanced costimulatory domains for improved T cell activation.
Co-Stim CAR Design Features optimized costimulatory signaling for enhanced T cell function.
Smarter™ Sonogenetic CART Development Service Significantly minimizing the risk of on-target, off-tumor toxicity, enhancing cytotoxicity and persistence.
TCR-controlled Fuzzy Logic CAR Construct Service Overcoming antigenic heterogeneity and escape, potentially improving persistence and function.
Multi-Target Strategies TriCAR Construction Targets three different antigens simultaneously for improved efficacy.
Compound CAR (cCAR) Construction Combines multiple CAR specificities in a single construct.
pan-ErbB-targeted Combination CAR-T Cell Targets multiple members of the ErbB family simultaneously.
These designs address specific challenges or applications in CAR-T cell therapy.
Categories Designs Features
Universal Platforms Smarter™ Universal ZipCAR Construction Provides a platform for rapid adaptation to different targets using zip-like connectors.
NKG2D-based Universal CAR Engineering Service Using TIM Molecule Combines NKG2D recognition with TIM molecules for broader applicability.
Synthetic T Cell Antigen Receptor (STAR) Construction Uses synthetic biology principles to create highly engineered receptor systems.
Special Applications PD-1-disrupted CAR Construction Includes PD-1 knockout for enhanced performance in immunosuppressive environments.
CRISPR/Cas9-based CAR Construction Uses CRISPR technology for precise genetic modifications in CAR-T cells.
CAR-Macrophage Adapts CAR technology for use in macrophages instead of T cells.
CAR-NK Vector Design and Construction Adapts CAR technology for use in Natural Killer cells.
Smarter™ Safety CAR Construction Incorporates multiple safety features for improved clinical safety.
CART Engineering Service Targeting TRBCs for T-cell Malignancies Immunotherapy Specifically designed for targeting T-cell malignancies.
CART Engineering Strategies for In Vivo Imaging Detection Includes imaging markers for tracking CAR-T cells in vivo.
Infographic Evolution of Chimeric Antigen Receptor Design This infographic presents the design of chimeric-antigen receptors (CARs), including:
  • CAR design and evolution
  • Comparison of different generations of CAR
  • Blueprint of CAR design

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Fig. 19 DIY your own chimeric antigen receptor design. (Creative Biolabs Original)
We have developed a proprietary online system for customizing CAR and CAR cell products, which offers full options to meet all unique needs, including but not limited to conventional or unconventional CAR constructs, as well as a variety of vectors and cells. The customization process can be completed with just a few simple clicks, please feel free to try it out.
DIY your own design

CAR Construction & Validation

1. Gene Sequence Design & Optimization

Starting with CAR gene sequence selection and codon optimization for efficient translation. Incorporates synthetic elements like promoters and signal peptides for optimal T cell expression.

2. Vector Selection

Evaluation of viral (lentiviral, adenoviral) and non-viral systems based on specific needs. Includes plasmid vectors and mRNA-based systems for reduced mutagenesis risk.

3. Cloning & Plasmid Construction

Multi-step cloning using restriction enzyme digestion and Gibson assembly for precise CAR gene integration. Focus on high-quality plasmid production.

4. Sequence Verification

Rigorous verification through Sanger and next-generation sequencing to confirm correct orientation and assembly of all components.

5. Expression Validation

Assessment of CAR expression via flow cytometry, Western blotting, and immunofluorescence to verify proper protein levels and localization.

6. Stability & Functional Testing

Extended culture period testing for CAR stability, cytotoxicity assays, and cytokine release evaluation to confirm therapeutic potential.



Why Choose Creative Biolabs

15+
Years Experience
2000+
Projects Delivered
100%
Client Satisfaction
Technical Expertise

Our team brings deep expertise in CAR-T therapy, developing both conventional and novel CAR formats. We leverage advanced techniques to ensure each CAR design maximizes therapeutic efficacy while minimizing potential risks.

Advanced Innovation

Creative Biolabs advances CAR development through innovative designs, including Tandem CARs, CRISPR/Cas9-based CARs, and Smarter™ self-destruct CARs, enabling more precise and safer therapies.

Rigorous Quality Control

Our verification and validation processes guarantee the accuracy, functionality, and stability of CAR constructs. This thorough approach ensures that you receive reliable, validated designs ready for downstream applications.

Streamlined Development Process

We accelerate research timelines through an integrated process combining CAR design, vector construction, and thorough testing. Our approach enables efficient progression from research to clinical applications.

Tailored Solutions

We provide customizable CAR design and vector construction options to meet your specific therapeutic goals. Our flexibility in design and optimization ensures solutions tailored to your research needs.

Tailored Solutions

Our experienced scientific team provides dedicated consultation throughout your project lifecycle. We offer strategic guidance and technical expertise backed by extensive practical experience in CAR-T development.



Trusted By Industry Leaders

OUR VALUED
PARTNERSHIPS

"What impressed us most about Creative Biolabs was not just their technical expertise, but the clarity and transparency in their communication. They kept us informed at every step, ensuring we understood the decisions being made and the potential impact on our project. We felt truly supported, and the CAR constructs we received were exactly what we needed for our next phase of research."

— Michael P., Director of Cell Therapy Research

"Working with Creative Biolabs was one of the most efficient experiences we’ve had in CAR development. From initial discussions to the final delivery of the CAR constructs, everything was handled with professionalism and care. Their turnaround times were impressive, and the quality of the constructs was impeccable. They understood our project's urgency and consistently delivered on time, without compromising on quality."

— Dr. Laura M., Head of Therapeutic Development

"Creative Biolabs played a critical role in helping us navigate the complexities of CAR-T development. Their team offered creative and practical solutions that were perfectly aligned with our research needs. Their strategic guidance and attention to detail transformed the way we approached CAR design. We achieved better outcomes faster than we had anticipated, and their support was pivotal throughout the process."

— Dr. Jane H., Principal Investigator, Immuno-Oncology

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.

Resources

Use the resources in our library to help you understand your options and make critical decisions for your study.

Engineering strategies for improving adoptive T cell therapy
Webinar: Engineering strategies for improving adoptive T cell therapy
Convertible CAR-T, a highly adaptable CAR-T platform
Webinar: Convertible CAR-T, a highly adaptable CAR-T platform
Chimeric antigen receptor CAR Technique
Chimeric antigen receptor CAR Technique
Advances in CAR-T Cell Therapy
Advances in CAR-T Cell Therapy
Understand CAR-T Therapy by One Graphic
Understand CAR-T Therapy by One Graphic
Evolution of Chimeric Antigen Receptor Design
Evolution of Chimeric Antigen Receptor Design
CAR-T Cell Immunotherapy: Development of CAR-T Cells
CAR-T Cell Immunotherapy: Development of CAR-T Cells
CAR-T Cell Immunotherapy: Clinical Applications and Challenges
CAR-T Cell Immunotherapy: Clinical Applications and Challenges
CAR-T Cell Immunotherapy: Development of CAR-T Cells
CAR-T Cell Immunotherapy: Basic Principles
CAR-T Cell Immunotherapy: Clinical Applications and Challenges
Obstacles and Potential Strategies in CAR-T Therapy
One-Stop CAR-T Therapy Development
One-Stop CAR-T Therapy Development
CAR-T Cell In Vitro Assay Services
CAR-T Cell In Vitro Assay Services
CAR Design & Construction Service
CAR Design & Construction Service
CAR-T Preclinical In Vivo Assay
CAR-T Preclinical In Vivo Assay

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CAR Therapy Development Services Portfolio

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