monoclonal antibodies

PART 1: INTRODUCTION TO MONOCLONAL ANTIBODIES

Section 1: Learning Outcomes (Page 2)

By the end of this lecture, students will understand:

• What's the market – the scale and growth of the monoclonal antibody (mAb) market.

• How they are made and naming conventions – production methods and the systematic nomenclature for mAbs.

• Formulation & reconstitution – practical aspects of handling and preparing mAb products for administration.

Much of the content is taken from: Lu, RM., Hwang, YC., Liu, IJ. et al. Development of therapeutic antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020).

PART 2: THE MONOCLONAL ANTIBODY MARKET

Section 2: The Growing Market (US FDA) (Page 3)

• A graph showing the increasing number of monoclonal antibody approvals by the US FDA over time.

• Trend: Significant growth, particularly from the mid-1990s onwards, with accelerating approvals in the 2010s and 2020s.

Section 3: The Global Market (Page 4)

Source: Precedence Research

Global Monoclonal Antibodies Market Size and Forecast:

• Key Drivers: Increasing prevalence of chronic diseases (cancer, autoimmune disorders), technological advancements, and expanding applications of mAbs.

Image Description (Page 4): A graph showing the projected growth of the global monoclonal antibodies market from 2023 to 2033.

PART 3: PRODUCTION AND NAMING OF MONOCLONAL ANTIBODIES

Section 4: How They Are Made (Page 5)

• The lecture notes that phage display technology is being avoided for today's discussion, focusing on other production methods.

• General Principle: Monoclonal antibodies are produced by hybridoma technology (fusion of B cells with myeloma cells) or recombinant DNA technology (expression in mammalian cells like CHO cells).

Section 5: Names from Murine vs. Human – The Naming Convention (Pages 6-7)

5.1. The Suffix "-mab" (Page 6):

• All monoclonal antibody names end with the suffix "-mab" .

5.2. Infixes Indicating Source/Target (Page 6):
The part of the name before "-mab" provides information about the antibody's origin and target.

Examples of Therapeutic Antibodies by Indication (Page 6):

5.3. WHO INN Nomenclature Scheme (2021) – New System (Page 7):

The World Health Organization (WHO) International Nonproprietary Names (INN) scheme for monoclonal antibodies was updated in 2021. The new system uses a different stem and infixes to indicate the target.

New Stem: -mab is replaced by -bart (?) – Note: The slide shows a complex table. The key change is a more systematic approach to naming based on target.

Substems (Pre-substem) and Infixes (Target):

Notes:

• At the 69th INN Consultation, the infix changed from -gros- to -gro- to avoid a conflict with the infix -os- .

• At the 70th INN Consultation, it was decided that antibodies targeting an interleukin receptor would also have the -ki- infix.

PART 4: FORMULATION AND EXCIPIENTS

Section 6: Two Examples from the SmPC (Page 8)

6.1. Remicade® (Infliximab) 100 mg powder for concentrate for solution for infusion:

6.2. Entyvio® (Vedolizumab) 300 mg powder for concentrate for solution for infusion:

Section 7: Antibody Structures (Page 9)

Image Description (Page 9): Three diagrams showing different antibody formats:

1. Traditional Monoclonal Antibody (IgG): Shows the classic Y-shaped structure with two heavy chains (blue) and two light chains (green). The Fab regions (antigen-binding fragments) and Fc region (crystallisable fragment) are labelled. Disulfide bonds are indicated.

2. Fab' Fragment: Shows a smaller fragment consisting of one light chain and part of one heavy chain, linked by a disulfide bond. This format lacks the Fc region.

3. PEGylated Fab' Fragment: Shows the Fab' fragment with a polyethylene glycol (PEG) chain attached. PEGylation increases the half-life of the fragment (which would otherwise be cleared rapidly). Example: Certolizumab pegol is a PEGylated Fab' fragment.

PART 5: SHELF LIFE AND STORAGE

Section 8: Infliximab (Remicade®) Storage (Page 10)

8.1. Before Reconstitution (Lyophilised Powder):

• Shelf life: 3 years at 2°C – 8°C (refrigerated).

• Flexible Storage: Remicade may be stored at temperatures up to a maximum of 25°C for a single period of up to 6 months, but not exceeding the original expiry date. The new expiry date must be written on the carton.

• Important: Upon removal from refrigerated storage, Remicade must not be returned to refrigerated storage.

8.2. After Reconstitution and Dilution:

• Chemical and physical in-use stability: Demonstrated for up to 28 days at 2°C to 8°C and for an additional 24 hours at 25°C after removal from refrigeration.

• Microbiological stability: From a microbiological point of view, the infusion solution should be administered immediately. In-use storage times and conditions prior to use are the responsibility of the user and would normally not be longer than 24 hours at 2°C to 8°C, unless reconstitution/dilution has taken place in controlled and validated aseptic conditions.

Section 9: Vedolizumab (Entyvio®) Storage (Page 10)

9.1. Shelf Life:

• 3 years (unopened, refrigerated).

9.2. In-Use Stability:

• Reconstituted solution in the vial: Demonstrated for 8 hours at 2°C – 8°C.

• Diluted solution in infusion bag (in sodium chloride 9 mg/mL): Demonstrated for:

  • 12 hours at 20°C – 25°C, OR

  • 24 hours at 2°C – 8°C.

• Combined in-use stability: The total combined time (vial + infusion bag) is:

  • 12 hours at 20°C – 25°C, OR

  • 24 hours at 2°C – 8°C.

  • A 24-hour period may include up to 8 hours at 2°C – 8°C for reconstituted solution in the vial and up to 12 hours at 20°C – 25°C for diluted solution in the infusion bag, but the infusion bag must be stored in the refrigerator (2°C – 8°C) for the rest of the 24-hour period.

• Do not freeze the reconstituted solution in the vial or the diluted solution in the infusion bag.

Storage Summary Table (Entyvio):

¹ Up to 30 minutes are allowed for reconstitution.
² This time assumes the reconstituted solution is immediately diluted and held in the infusion bag only. Any time the reconstituted solution was held in the vial should be subtracted.
³ This period may include up to 12 hours at 20°C-25°C.

PART 6: RECONSTITUTION AND HANDLING

Section 10: Reconstitution Instructions – Remicade® (Infliximab) (Page 11)

1. Use aseptic technique.

2. Reconstitute each vial with 10 mL of Water for Injections, using a syringe with a 21-gauge (0.8 mm) or smaller needle.

   • Remove flip-top, wipe with 70% alcohol swab.

   • Insert needle through centre of rubber stopper.

   • Direct the stream of water to the glass wall of the vial (to minimise foaming).

3. Gently swirl the solution by rotating the vial to dissolve the lyophilised powder.

   • Avoid prolonged or vigorous agitation.

   • DO NOT SHAKE.

   • Foaming is not unusual.

4. Allow the reconstituted solution to stand for 5 minutes.

5. Check the solution: It should be colourless to light yellow and opalescent. The solution may develop a few fine translucent particles (as infliximab is a protein).

   • Do not use if opaque particles, discolouration, or other foreign particles are present.

Section 11: Reconstitution Instructions – Entyvio® (Vedolizumab) (Page 11)

1. Use aseptic technique.

2. Remove flip-off cap, wipe with alcohol swab.

3. Reconstitute vedolizumab with 4.8 mL of sterile Water for Injections at room temperature (20°C-25°C) , using a syringe with a 21-25 gauge needle.

4. Insert needle through centre of stopper, direct stream of liquid to the wall of the vial to avoid excessive foaming.

5. Gently swirl the vial for at least 15 seconds. Do not vigorously shake or invert.

6. Let the vial sit for up to 20 minutes at room temperature to allow for reconstitution and for any foam to settle. The vial can be swirled and inspected during this time. If not fully dissolved after 20 minutes, allow another 10 minutes.

7. Inspect: Solution should be clear or opalescent, colourless to light yellow, and free of visible particulates. Do not use if uncharacteristic colour or particulates are present.

8. Once dissolved, gently invert vial 3 times.

Section 12: Dilution and Administration (Page 12)

12.1. Remicade® (Infliximab) Dilution:

1. Dilute the total volume of reconstituted Remicade solution to 250 mL with sodium chloride 9 mg/mL (0.9%) solution for infusion.

   • Do not dilute with any other diluent.

   • Withdraw a volume of saline from the 250 mL bag equal to the volume of reconstituted Remicade.

   • Slowly add the total volume of reconstituted Remicade to the infusion bag.

   • Gently mix.

2. For volumes greater than 250 mL, use a larger infusion bag (e.g., 500 mL, 1000 mL) or multiple 250 mL bags to ensure the concentration does not exceed 4 mg/mL.

3. If stored refrigerated after reconstitution and dilution, the infusion solution must be allowed to equilibrate at room temperature to 25°C for 3 hours prior to infusion.

4. Administer over the recommended infusion time (see SmPC section 4.2).

   • Use an infusion set with an in-line, sterile, non-pyrogenic, low protein-binding filter (pore size 1.2 micrometre or less) .

5. Start administration as soon as possible and within 3 hours of reconstitution and dilution (if not stored refrigerated). If refrigerated, the 24-hour rule applies (see Section 8.2).

6. Do not infuse Remicade concomitantly in the same intravenous line with other agents.

7. Visually inspect for particulate matter or discolouration prior to administration. Do not use if visible particles are observed.

8. Dispose of any unused product according to local requirements.

12.2. Entyvio® (Vedolizumab) Dilution:

1. Immediately withdraw 5 mL (300 mg) of reconstituted Entyvio using a syringe with a 21-25 gauge needle.

2. Add the 5 mL of reconstituted Entyvio to 250 mL of sterile sodium chloride 9 mg/mL (0.9%) solution for injection.

   • Gently mix the infusion bag.

   • (5 mL of saline does NOT need to be withdrawn from the bag prior to adding Entyvio).

3. Do not add other medicinal products to the prepared infusion solution or intravenous infusion set.

4. Administer the infusion solution over 30 minutes.

5. Once reconstituted, the infusion solution should be used as soon as possible.

6. Do not store any unused portion for reuse. Each vial is for single use only.

7. Dispose of unused product according to local requirements.

PART 7: KEY POINTS SUMMARY