Monoclonal Antibodies

Learning Outcomes

  • Understanding the market for monoclonal antibodies.

  • Exploring the production methods of monoclonal antibodies.

  • Learning about naming conventions regarding monoclonal antibodies.

  • Familiarizing with the formulation and reconstitution processes of monoclonal antibodies.

  • Source Material: Development of therapeutic antibodies for the treatment of diseases. Lu, RM., Hwang, YC., Liu, IJ. et al. J Biomed Sci 27, 1 (2020).

The Market for Monoclonal Antibodies

  • The market for monoclonal antibodies is projected to grow significantly:

    • Estimated market value in 2025: $300 billion

    • Notable mAbs and their therapeutic uses:

    • Caplacizumab (Cablivi): anti-vWF for acquired Thrombotic Thrombocytopenic Purpura (aTTP)

    • Romosozumab (Evenity): anti-Sclerostin for osteoporosis treatment

    • Burosumab (Crysvita): anti-FGF23 for X-linked Hypophosphatemic Rickets (XLH)

    • Ibalizumab (Trogarzo): anti-CD4 for HIV infection

    • Erenumab (Aimovig): anti-CGRPR for migraine prevention

    • Rituximab (Rituxan): anti-CD20 for Non-Hodgkin lymphoma

    • Daclizumab (Zinbryta): anti-CD25 for multiple sclerosis

    • Atezolizumab (Tecentriq): anti-PD-L1 for bladder cancer

    • Reslizumab (Cinqaero, Cinqair): anti-IL-5 for asthma

  • Historical Data: Market value development over time

    • 1975-2025 market evaluation shows increasing investment and value developments, e.g.:

    • From $0.3 billion in 1997 to projected $300 billion in 2025.

Production of Monoclonal Antibodies

  • Common methods of producing monoclonal antibodies:

    • (A) Mouse Hybridoma:

    • B cells from the spleen of immunized mice are fused with myeloma cells.

    • (B) Phage Display:

    • Libraries of phages displaying antibodies are utilized to find binding candidates.

    • (C) Transgenic Mouse:

    • Mice genetically modified to express human antibodies.

    • (D) Single B Cell Sorting:

    • Isolated B cells from peripheral blood mononuclear cells (PBMC) are sorted for specific binding.

    • Following isolation, antibodies are screened using biopanning with antigens.

  • Humanization Techniques:

    • Mouse mAb development is generally followed by chimerization and CDR grafting to create chimeric mAbs (mouse and human components).

    • Final step constructs decide the structure of human Immunoglobulin (IgG).

Naming Conventions

  • Monoclonal antibodies are named using a systematic nomenclature established by the World Health Organization (WHO):

    • Infix Definitions:

    • -ami- for serum amyloid protein (SAP)/amyloidosis.

    • -ba- for bacterial.

    • -ci- for cardiovascular medications.

    • -de- for metabolic or endocrine pathways.

    • -eni- for enzyme inhibition.

    • -fung- for fungal targets.

    • -gro- for cytokines and receptors.

    • -ki- for allergens.

    • -ler- for immunosuppressive uses.

    • -ne- for nervous system applications.

    • -os- for bone-related treatments.

    • -ta- for tumor targeting.

    • -toxa- for toxins.

    • -vet- for veterinary applications.

    • -vi- for viral diseases.

    • Changes in nomenclature over the years:

    • Change from –gros- to –gro- to avoid conflicts.

    • Inclusion of -ki- infix for interleukin receptor targeting antibodies.

Formulation & Reconstitution

  • Formulation Examples:

    • Remicade® (infliximab):

    • Form: 100 mg powder for concentrate for solution for infusion.

    • Excipients: Sucrose, Polysorbate 80, Monobasic sodium phosphate, Dibasic sodium phosphate.

    • Entyvio (vedolizumab):

    • Form: 300 mg powder for concentrate for solution for infusion.

    • Excipients: L-histidine, L-histidine monohydrochloride, L-arginine hydrochloride, Sucrose, Polysorbate 80.

  • Shelf Life and Storage:

    • Infliximab:

    • Before reconstitution: 3 years at 2°C–8°C.

    • After reconstitution: Demonstrated stability for up to 28 days at 2°C–8°C, and an additional 24 hours at 25°C.

    • Vedolizumab:

    • Reconstituted: stability for 8 hours at room temperature, 12 hours if diluted in saline.

Special Handling Procedures

  • Reconstitution Techniques (Remicade):

    1. Under aseptic conditions, reconstitute each Remicade vial with 10 ml of water for injections using a 21-gauge needle or smaller.

    2. Gently swirl the vial to dissolve the lyophilized powder, avoid vigorous agitation or shaking.

    3. After reconstitution, allow the solution to stand for 5 minutes.

    4. Inspect for coloration and particle presence before administration; a clear or opalescent solution is preferred.

  • Dilution for Administration (Remicade):

    • Dilute reconstituted solution to 250 ml using sodium chloride 9 mg/ml (0.9%) solution.

    • Administer without preservatives as soon as possible, ideally within 3 hours post-dilution.

Conclusion

  • Comprehensive understanding of monoclonal antibodies involves knowledge of the market, production processes, naming conventions, as well as formulation and handling practices. These elements are crucial for ensuring the effective use of monoclonal antibodies in clinical settings.

References

  • Various URLs and documents were referenced during the lectures for further information on monoclonal antibodies, including:

    • Scientific articles, summaries of product characteristics (SmPC), and other academic resources related to therapeutic monoclonal antibodies.