Monoclonal Antibody Production Training Program

Build job-ready skills for end-to-end monoclonal antibody production. This program covers antigen design, animal immunization, PEG-mediated fusion, HAT selection, screening and subcloning, culture scale-up, Protein A/G purification, polishing, QC analytics, banking and tech transfer documentation.

Module 1 – Fundamentals of Monoclonal Antibodies (Theory - Online)

1. Antibody basics: domains, isotypes, affinity vs avidity, epitope concepts.
2. mAb landscape: hybridoma route vs recombinant discovery, typical use-cases.
3. Antigen strategies: peptide, protein, conjugates; carrier and hapten logic.
4. Adjuvants overview and immunization schedules; humane and ethical practices.
5. Hybridoma principle: fusion partners, HAT selection, clonality rationale.
6. Screening funnels: ELISA, specificity, cross-reactivity, isotyping basics.
7. Scale-up paths: static flasks, spinner, wave; serum-free adaptation overview.
8. Purification options: Protein A/G/L, precipitation and polishing outlines.
9. QC and stability concepts: purity, aggregation, endotoxin and mycoplasma awareness.
10. Project documentation mindset: SOPs, batch records and data integrity.

Module 2 – Antigen Design & Preparation (Practical)

1. Antigen selection criteria; peptide vs full-length protein decision tree.
2. Expression and purification overview for recombinant antigens; tag choices.
3. Conjugation to carriers (e.g., KLH, BSA) for hapten and small protein targets.
4. Buffer, pH and storage optimization; stabilizers and preservative choices.
5. Endotoxin awareness and reduction workflow outlines.
6. Analytical QC: SDS-PAGE, Western, UV280 and concentration checks.
7. Aggregation checks by simple SEC/filtration and visual inspection.
8. Aliquoting, freeze-thaw policy and chain-of-custody records.
9. Plate-coating optimization for ELISA screening readiness.
10. Risk log and acceptance criteria definition for proceeding to immunization.

Module 3 – Immunization & Titer Monitoring (Practical)

1. Species and strain selection; cohort sizing and randomization basics.
2. Adjuvant emulsions and injection routes; schedule planning.
3. Bleed schedule, serum processing and storage conditions.
4. Indirect ELISA titering; cutoff definition and serial dilution curves.
5. Specificity and cross-reactivity panel design; negative and positive controls.
6. Boosting decisions based on titer kinetics and quality of response.
7. Recordkeeping templates and labeling standards.
8. Animal welfare, biosafety practices and waste handling.
9. Pre-fusion readiness checklist and donor selection.
10. Shipment/transfer logistics to fusion lab when applicable.

Module 4 – PEG-Mediated Fusion & Hybridoma Generation (Practical)

1. Spleen harvest and splenocyte preparation; viability and counts.
2. Myeloma line revival and culture; log-phase readiness and selection markers.
3. PEG-mediated fusion workflow; ratios, timing and gentle handling.
4. Post-fusion handling; dilution, plating densities and feeder cells.
5. HAT/HT selection setup; media prep and exchange schedules.
6. Incubation and contamination avoidance practices.
7. Initial microplate monitoring and clone emergence tracking.
8. Early screening plate map design and sample tracking.
9. Basic troubleshooting of low fusion efficiency.
10. Safety and aseptic technique reinforcement.

Module 5 – Screening, Subcloning & Clonality (Practical)

1. Primary ELISA screening; cutoff setting and hit triage rules.
2. Confirmatory assays; counter-screening vs close homologs.
3. Isotyping of promising clones; kappa vs lambda chains awareness.
4. Limiting dilution subcloning; Poisson logic and plate layouts.
5. Clonality verification over passages; stability checks.
6. Early cryopreservation of hit clones to mitigate loss.
7. Mycoplasma testing workflow and remediation options.
8. Small-scale production to confirm secretion levels.
9. Ranking matrix: potency, specificity and growth traits.
10. Documentation of clone histories and change logs.

Module 6 – Hybridoma Expansion & Antibody Harvest (Practical)

1. Scale-up strategies: static flasks, spinner and basic perfusion outlines.
2. Serum-free and protein-free media adaptation pathways.
3. Feeding and viability monitoring; key culture KPIs.
4. Conditioned medium collection and clarification.
5. Pre-capture assessments: titer estimate and binding to A/G screens.
6. Precipitation alternatives (ammonium sulfate, caprylic acid) awareness.
7. Depth filtration and bioburden control.
8. Hold conditions and cold chain for harvest pools.
9. Basic yield projections and mass balance notes.
10. Batch record creation for each harvest.

Module 7 – Purification & Polishing (Practical)

1. Protein A/G/L capture selection based on isotype and subclass.
2. Binding, wash and elution optimization; neutralization strategy.
3. Buffer exchange and diafiltration (spin/TFF) to target formulation.
4. SEC polishing for monomer percentage; aggregation control.
5. Endotoxin awareness and removal workflows.
6. Analytical QC: SDS-PAGE, UV280, purity estimates.
7. Optional precipitation cleanup and re-capture tactics.
8. Sterile filtration and fill-finish practices for lab scale.
9. Storage stability conditions and label content.
10. Deviation and CAPA notes for run issues.

Module 8 – Characterization & Validation (Practical)

1. Binding kinetics by BLI/SPR (overview) and apparent KD estimation.
2. Epitope mapping basics and competitive ELISA/binning matrix.
3. Functional assay outlines (neutralization or blocking where relevant) .
4. Specificity panel vs related antigens; off-target checks.
5. Thermal stability (DSF/Tm) and stress-challenge quick screens.
6. Basic glyco-awareness for mouse mAbs; Fc-related notes.
7. Endotoxin and HCP awareness assays (workflow) .
8. Acceptance criteria and report templates.
9. Data integrity and raw data archiving.
10. Readiness gate to bank and scale-up.

Module 9 – Banking, Cryopreservation & QC (Practical)

1. Freeze medium composition and controlled-rate freezing.
2. Master and working cell bank plan; vial counts and mapping.
3. Post-thaw recovery testing and secretion re-checks.
4. Identity checks (isotype profile) and sterility/mycoplasma testing.
5. Viability and passage-limit policy for production.
6. Duplicate storage and contingency planning.
7. Inventory, labels and LIMS basics.
8. Audit trail expectations and documentation.
9. Transport and dry shipper handling notes.
10. Periodic re-qualification schedule.

Module 10 – Tech Transfer, Compliance & Capstone (Practical)

1. SOP pack assembly: methods, acceptance criteria and templates.
2. Batch records and run summaries; deviations and CAPA notes.
3. QC release checklist: identity, purity and endotoxin awareness.
4. Basic regulatory and ethics touchpoints for preclinical mAbs.
5. Tech transfer dossier and receiving-lab checklist.
6. Costing, timelines and resource planning for a pilot build.
7. Troubleshooting playbook for low titer, contamination and aggregation.
8. Risk registry and mitigation planning.
9. Capstone mini-project presentation with data pack.
10. Career portfolio curation and next steps.