Bioconjugation Services

Adding a small molecule to a protein might seem simple, but the process is ridden with challenges. It requires advanced analytical instruments, specialized purification equipment, and expertise in both chemical and biological handling that demands significant investment in infrastructure and skilled resources. NJ Bio is well equipped with the required analytical and purification instruments, chemical and biological handling requirements, and human resources, with a deep understanding and experience of chemical biology, synthetic chemistry, and biochemistry. NJ Bio offers solutions in those areas that complement our clients’ expertise and build a lasting relationship based on quality work, transparency, and adding value to our clients’ drug development programs. We work proactively to materialize the bioconjugation product for the intended application.

Affordable, Customizable and Expert Solutions

NJ Bio has strong bioconjugation expertise in various proteins, linker payloads, and conjugation technologies. We offer different service options to maximize value for our clients and perform bioconjugations at a stage appropriate level. Our purified conjugates are accompanied by robust analytics to ensure products are well-characterized so that you can understand and interpret the data generated on purity, DAR and residual analysis. We can assist in selecting suitable linker-payload combinations tailored to the biology of your target. With a proven track record in managing complex bioconjugation projects, our team is highly skilled in recommending the optimal conjugation technology to achieve your study’s desired DAR, monomeric purity, and other desirable attributes. We deliver these services efficiently through our integrated CRDMO platform, enabling seamless transition, cost management and accelerated time-to-market.

Discovery Service Options

* Timelines may vary depending on the scale of the project

Bioconjugation services involve the precise and controlled coupling of biomolecules, such as antibodies, proteins, enzymes, or other bioactive compounds, with various chemical entities, nanoparticles, or surfaces. Bioconjugation experts leverage their expertise to design, optimize, and perform custom conjugation reactions, resulting in tailored bioconjugates with enhanced functionality and stability. These services are essential for applications in antibody drug conjugates, diagnostics, drug development, targeted therapies, and research, facilitating the creation of novel tools, assays, and therapeutic agents. NJ Bio’s skilled team of scientists can ensure the successful integration of biological and chemical components of a drug bioconjugates, achieving specific objectives of clients’ research and development projects. With a focus on bioconjugation, we deliver comprehensive solutions that support your ADC programs from preclinical research through to early-stage clinical manufacturing. Our specialized ADC services encompass every stage of ADC development, from custom antibody production and payload-linker synthesis to bioconjugation and manufacturing.

Express Conjugation: Early discovery and proof-of-concept (only for ADCs)

NJ Bio’s Express Conjugation Service is an expeditious and cost-effective platform to assess an ADC target using validated linker-payloads and stochastic lysine- or cysteine-based conjugation. Our team carries out characterization of ADC using SEC, MS, UV and HPLC to ensure the quality of the final product. Since the antibody doesn’t need to be humanized at this stage, we can conjugate a variety of IgG species and isotypes to determine the best option for further development studies.

Example: You have 6 murine antibody candidates and want to screen them in vitro with 1 or 2 approved linker-payloads (such as SMCC-DM1, MC-Val-Cit-PAB-MMAE, MC-MMAF, DXd(1) or CL2A-SN38) to help with your selection criteria. NJ Bio can rapidly help you study these leads in vitro and preliminarily in vivo to justify whether to move to the next stage.

Standard Conjugation

Our standard conjugation service is a general conjugation platform that helps you understand the MoA, release mechanism and potency range required for your target. With access to a broader range of validated linker-payloads (e.g., DGN549, Tesirine, DM21C, or SPDB-DM4), this platform will provide material for in vivo selection. At this stage, we recommend utilizing validated payloads with stochastic conjugation, however clients may also explore site-specific conjugation technologies based on their project requirements.

Example: You have proof of concept with MC-VC-MMAE (Vedotin) that shows in vitro activity and some activity in vivo. You would like to assess payloads with different mechanisms of action such as topoisomerases or DNA damaging agent(s) in a larger in vivo study with a dose response. The Standard Conjugation option can help ascertain whether you will have a clinical candidate with an optimized ADC.

Custom Conjugation

The Custom Conjugation platform allows you to determine the best bioconjugate for preclinical development and lead optimization. Validated payloads do not always have the best therapeutic index (TI) for a particular indication. Our scientists can mix and match different payloads, linker technologies, and site-specific technologies to obtain the most appropriate candidate within your time constraints. This service allows to maximize stability, increase hydrophilicity, and ensure proper PK and release kinetics. During lead optimization, larger quantities of material are needed for toxicology studies, which also requires larger quantities of linker-payloads. Through our flexible FTE program, resources are allocated as needed to efficiently provide high-quality material to accelerate your project. NJ Bio can also develop a selected conjugate/ ADC based on the clients’ project requirements.

Example: You have an ADC that shows tumor regression, but the TI is narrow. To maximize clinical success, an improvement to the TI is beneficial. This can be done by testing different site-specific technologies, increasing the hydrophilicity of the linker-payload, and ensuring the ADC is optimized for the expected clinical dose range.

Process Development Services

We offer comprehensive support in optimization, and scale-up of bioconjugates to meet the demands of both preclinical and clinical development. We have years of experience in optimizing processes through Design of Experiments (DoE) and producing high-quality materials at scale for preclinical studies. Our in-house capabilities for scaling up linker-payload chemistry ensure a seamless technical transfer when moving to clinical manufacturing. The processes we develop for conjugation and linker-payload synthesis are robust, reproducible, and tailored for efficient transfers, minimizing risks and ensuring consistency across production stages.

In addition, our analytical and bioanalytical teams are equipped to develop and validate methods throughout the drug development lifecycle. From structural characterization and purity assessment to stability studies and potency assays, we provide the critical data needed to advance your bioconjugate project with confidence.

Clinical Material Manufacturing

GMP bioconjugation involves producing bioconjugates under controlled GMP conditions to ensure product consistency, safety, efficacy, and compliance with regulatory standards. At our cGMP suite in Princeton, we support small-scale bioconjugation projects, with plans to further expand manufacturing capabilities in 2025. Our dedicated Small Molecules Chemistry Suite enables the production of Phase 1 and Phase 2 clinical trial materials, such as linker payloads, in a more efficient and cost-effective manner compared to larger commercial suppliers. Our goal is to have a seamless transition across method development, engineering runs, and production of key clinical materials.  We also offer GMP Analytical Services for characterization, potency testing, and stability studies tailored to customer requirements. We provide end-to-end support for bioconjugate development, from initial feasibility studies to process and analytical method development, to the manufacturing of bulk drug substance (DS) for first-in-man studies.

Diverse Bioconjugation Technologies:

The choice of bioconjugation strategy depends on factors such as the target antigen, payload, required stability, and desired DAR. Our bioconjugation experts, with years of experience, provide guidance in selecting the optimal strategy for your project. NJ Bio offers a wide range of bioconjugation methods, including lysine- and cysteine-based modifications, as well as advanced site-specific techniques such as affinity-based conjugation, oxime and biorthogonal chemistries, glycan remodeling, transglutaminase-based methods, non-natural amino acid incorporation, click chemistry, bifunctional linker approaches and enzyme-assisted conjugation. With our expertise and state-of-the-art methods, we ensure the development of scalable and robust bioconjugation processes optimized for your study.

Case Study: Transglutaminase-mediated azide functionalization of mAb

To produce consistent and stable ADCs, precise site-specific functionalization is essential. Standard deglycosylation protocol using PNGase, however, led to partial deglycosylation. To address this, we used a modified deglycosylation protocol and implemented a transglutaminase (TG)-mediated method for introducing azide groups into mAbs at specific sites, enabling controlled conjugation of DBCO payloads. Complete deglycosylation via PNGase treatment successfully removed all N-linked glycans from the mAb, as confirmed by mass spectrometry (MS), which revealed a single peak for the deglycosylated species. TG catalyzed the site-specific attachment of azide-functionalized PEG derivatives to the mAb. This selective modification preserved the antibody’s structural integrity and antigen-binding properties. MS analysis confirmed complete deglycosylation, uniform azido-functionalized mAbs, and a defined DAR, with distinct peaks for conjugated species.

Common Bioconjugation Requests

We at NJ Bio have vast experience in the conjugation of different linker-payload combinations. Beyond cytotoxins like auristatin, maytansine, PBD and deruxtecan we have achieved conjugation of non-toxic functionalities such as peptides, oligonucleotides and other targeted therapeutic platforms.

Protein and Peptide Drug Conjugates

Peptide-drug conjugates (PDCs) combine peptides with drug molecules, requiring specialized synthesis and testing due to their complex structures. Given the great diversity of structures in proteins and peptides, the conjugation technologies developed for antibodies (e.g., cysteine, lysine conjugation) are not always applicable to all proteins. NJ Bio can adapt ADC technology for use with non-antibody proteins. For smaller proteins and peptides, the linker position, drug-linker properties, and DAR are critical parameters to consider. Our analytical techniques and equipment are fully adaptable to smaller conjugates. We have made substantial investments in mass spectrometry expertise and equipment, which are particularly effective for characterizing smaller conjugates. By leveraging our deep expertise in conjugation chemistry, we design these molecules to deliver high-quality bioconjugates with enhanced properties.

Carrier Protein Conjugates

NJ Bio has experience in the attachment of proteins to carrier agents that elicit an immune response. Common carrier agents include BSA (bovine serum albumin), KLH (keyhole limpet hemocyanin) and OVA (ovalbumin). These carriers are often designed to generate an antibody response to a linked antigen and are important tools in antibody discovery. We have experience in the conjugation of proteins to carrier agents, with applications from vaccines to antibody generation.

Oligonucleotide Conjugates

Antibody-oligonucleotide conjugates (AOCs) are a promising targeted drug delivery approach, combining oligonucleotides like siRNAs or ASOs with antibodies or fragments. AOCs offer advantages such as increased specificity, reduced off-target effects, and improved pharmacokinetics. NJ Bio specializes in custom AOC synthesis using proven coupling strategies, with expertise in cross-linking chemistry for efficient oligonucleotide conjugation. Successful AOC development requires careful attention to antibody selection, conjugation chemistry, and antigen internalization. Not all linkers compatible with small molecules work for oligonucleotides, and linker position and type impact conjugation efficiency. Our investment in mass spectrometry aids AOC optimization. NJ Bio has also published a review titled – Antibody-Oligonucleotide Conjugates: A Twist to Antibody-Drug Conjugations, summarizing key advancements and challenges in AOC therapeutics.

Other Conjugates (e.g., fluorescent dye-labelled, biotinylated, derivatized with chelators)

At NJ Bio, we have experience in the labelling of proteins for other applications. Conjugation of dyes that allow tracking by fluorescence or by microscopy is routinely performed. Biotinylation is an important tool for the capture of specific proteins for analysis by ELISA and mass-spectrometry. Our chemistry experience is an asset when a linker-dye combination is not commercially available, allowing us to perform the synthesis and conjugation under one roof.

NJ Bio offers the following bioconjugation services:

• Rapid ADC Validation and Proof-of-Concept (PoC)
• Early De-risking and Optimization of conjugation strategies
• Process Development and Optimization for scalable manufacturing
• Comprehensive Analytical Characterization of Bioconjugates: Physical, chemical, and spectrometry analysis, Primary structure determination
• Analytical Method Development, Release and stability testing
• Small-Scale GMP Bioconjugation for clinical supply
• Flexible Conjugation Platforms: Native and site-specific conjugation
• Protein- and peptide-drug conjugation
• AOC Synthesis, Analysis, and Purification
• Biological Characterization of Bioconjugates: Efficacy, binding, and safety assessments

Frequently Asked Questions

Can you customize payload-linker synthesis to meet specific ADC requirements?

Yes, we specialize in tailoring payload-linker synthesis based on your project needs. Our expertise includes optimizing synthetic routes, ensuring the highest purity and stability, and scaling the synthesis to meet preclinical and clinical batch requirements. To know more visit https://njbio.com/payloads-linkers-antibody-drug-conjugates/

How do you ensure batch-to-batch consistency for ADCs?

We maintain stringent process control and use advanced analytical techniques such as HIC-HPLC, SEC, LC-MS, and CE-SDS to assess critical quality attributes, including DAR, purity, aggregation, and stability.

What analytical and bioanalytical services do you provide for ADC characterization?

Our capabilities include – Determination of DAR and conjugation efficiency, Purity profiling (HPLC, SEC, CE-SDS), Stability studies under regulatory guidelines, Potency testing (e.g., cytotoxicity assays), Residual solvent and endotoxin testing, and much more. To know more visit https://njbio.com/purification-and-analytical-services/

Can you assist in GMP manufacturing?

Yes, we can support small-scale GMP bioconjugation projects and provide robust payload-linker capabilities to support early-phase clinical trials, including Phases I and II. To know more visit https://njbio.com/manufacturing-services/