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Antibody Production

Reproducible and Time-efficient Services

Monoclonal antibodies (mAbs) are laboratory-engineered molecules designed to specifically bind to target antigens, mimicking the immune system’s attack on unwanted cells, such as cancer cells. mAbs are versatile tools used either on their own as therapeutic agents or biomarkers, or as carriers for delivering drugs or toxins directly to cancer cells through antibody-drug conjugates (ADCs). At NJ Bio, we leverage years of expertise in bioconjugation to produce monoclonal antibodies tailored for various research applications.

Our custom monoclonal antibody platform integrates seamlessly into larger bioconjugation projects, producing both standard and engineered mAbs based on client requirements. These antibodies are generally produced in CHO cell lines and purified from the supernatant, characterized, and tested to meet quality standards and project requirements. A standard project results in purified mAbs ready in less than twelve weeks from initiation, delivering timely and reliable solutions for your research projects.

Finding the right monoclonal antibody from thousands of options is just the first challenge. Ensuring successful synthesis, purification, and consistent, reproducible results are equally critical. Our technical staff has extensive experience in microbiology, cell biology, and bioconjugation, and many years of expertise in project design and planning ensuring that your projects are handled professionally and efficiently. Our custom monoclonal antibody generation service delivers highly specific, quality products with high affinity, designed to support scientific discovery with the reliability and precision you need.

Comprehensive Custom Antibody Solutions

Custom Monoclonal Antibodies: We offer custom antibody production services, generating standard and engineered mAbs based on client’s project requirements. Our platform integrates seamlessly into larger bioconjugation projects.

Variable Region Integration: Utilize our constant region framework sequence to integrate with your variable region sequence or provide us with the entire antibody sequence for production.

Antibody Validation Approaches: Utilizing a robust analytical platform for various cell-free assays such as BLI and ELISA to measure binding kinetics of antibody-antigen interactions, flow cytometry for interaction studies, IncuCyte® for internalization assessments, and plate-based IC50 analysis for antibody-drug conjugates (ADCs), etc.

Development Process:

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At NJ Bio, we guarantee the timely delivery of high-quality mAbs, with a standard turnaround time of under 12 weeks. If you provide the plasmids, we can expedite the process to just 4–6 weeks. Should you require us to create DNA constructs from amino acid sequences, please allow an additional 4 weeks. Our expertise in mammalian expression systems ensures production of high-quality therapeutic antibodies, primarily produced in mammalian host cell lines like ExpiCHO cells (Chinese Hamster Ovary cells).¹ CHO cells are responsible for approximately 70% of recombinant protein production today due to their high productivity and acceptable product quality attributes.² Cells are typically grown in flasks on a shaker platform in a humidified CO₂ cell culture incubator.

After harvesting the supernatant, we subject it to rigorous antibody purification and characterization processes. Our comprehensive downstream processes include purification, isolation of fragments, titer determination, and conjugation. We can purify mg-to-gram scale quantities of mAbs using a wide array of purification techniques. Protein-A affinity chromatography is our preferred method for high-yield IgG antibody purification, as it offers high selectivity and purity by directly loading cell culture supernatants. This method effectively removes host cell proteins (HCP), host DNA, process-related impurities, and potential viral contaminants, easing the burden on subsequent polishing steps necessary to meet quality requirements.³

For further characterization and quality control, we utilize Gel Electrophoresis (SDS-PAGE), and LC-MS. SDS-PAGE provides insights into protein size (molecular weight) and yield (quantity).

Fig 1: Monoclonal antibody characterization, A. SDS-PAGE analysis of mAb; 2 µg of mAb run either under reduced or non-reduced condition on 4-12 % Bis-Tris gel, B. RP-LC/MS0; 5 µg of de-glycosylated mAb injected over PLRP-S (1000 Å, 8 μm, 2.1 x 50 mm). C. Size exclusion chromatography: 20 µg mAb injected over TSKgel QPAK GFC 300 (0.5 mL/min, 1x PBS w/ 10 % IPA)

LC-MS enables comprehensive characterization of antibody-based therapeutics. This includes molecular weight determination, identification of modifications, antibody-drug conjugate analysis, stability testing, PK/PD studies, and higher-order structure analysis.⁴

Fig 2: Applications of LC-MS for the biophysical characterization of antibody-based therapeutics⁴

Our purified antibodies are also tested in cell-based assays such as IC50, internalization, binding to cells, and ELISA. For more details about these assays, visit https://njbio.com/cell-based-functional-assay-including-cytotoxicity-assays/

NJ Bio provides comprehensive monoclonal antibody (mAb) services:

Custom mAb production and antibody engineering
Highly specific, high-affinity monoclonal antibodies
Rapid production times, delivering mAbs in <4–6 weeks*
In vitro production using mammalian cell lines viz. ExpiCHO
Scalable production from 50 mg > 1 gram**
A team of dedicated, highly experienced technical experts to design and manage your project
Comprehensive analytical platform including cell-free assays like BLI/ ELISA to measure binding kinetics of antibody-antigen interactions, flow cytometry-based interaction, IncuCyte^® based internalization and plate based IC50 analysis of ADCs

*Timeline applies if plasmid is provided. If only the amino acid sequence is provided, an additional 4 weeks will be required.

** Depends on expression levels

References

(1) Li, F., Vijayasankaran, N., Shen, A., Kiss, R., & Amanullah, A. (2010, September). Cell culture processes for monoclonal antibody production. In Mabs, Vol. 2, No. 5, pp. 466-479. Taylor & Francis.

(2) Jayapal, K. P., Wlaschin, K. F., Hu, W., & Yap, M. G. (2007). Recombinant protein therapeutics from CHO cells-20 years and counting. Chemical engineering progress, 103(10), 40.

(3) Pinto, I. F., Aires-Barros, M. R., & Azevedo, A. M. (2015). Multimodal chromatography: debottlenecking the downstream processing of monoclonal antibodies. Pharmaceutical Bioprocessing, 3(3), 263-279.

(4) Robotham, A. C., & Kelly, J. F. (2020). LC-MS characterization of antibody-based therapeutics: recent highlights and future prospects. In Approaches to the Purification, Analysis and Characterization of Antibody-Based Therapeutics (pp. 1-33). Elsevier.

Frequently Asked Questions (FAQs)

What information do I need to provide for custom in vitro antibody production?

Clients need to provide the desired antibody protein sequence for both light and heavy chain.

What are the steps involved in antibody production?

Steps typically include optimization of codon for antibody sequences*, cloning of gene into plasmid*, transfection into host cells, cell culture, antibody purification, characterization and quality control.

*We currently outsource the codon optimization for antibody sequences and the gene cloning into the plasmid. These processes may require a three-way Confidential Disclosure Agreement (CDA) to ensure confidentiality.

How do you ensure the specificity of the produced antibodies?

The specificity of the produced monoclonal antibodies can be ensured through various in-house cell-based functional assays. These include binding assays with purified antigens or cells, employing techniques such as ELISA, BLI, or flow cytometry.

Can you assist with antibody conjugation or labeling?

Yes, we offer services such as antibody conjugation to small molecules, drugs, enzymes, etc., for various research applications.