Biomanufacture of Monoclonal Antibodies

1. This unit of the NBC2 global biomanufacturing curriculum provides curricular material  for producing monoclonal antibody biotherapeutics. Directive documents are listed for mAb production from NISTCHO cells and CHO-DP12 cells.

PRODUCTION OF cNISTmAb FROM NISTCHO CELLS

NISTCHO is a recombinant Chinese hamster ovary cell line expressing cNISTmAb that was developed by NIST and MilliporeSigma for use as a reference material. The following journal article describes the creation and characterization of the cell line.

Development and Characterization of the NISTCHO Reference Cell Line - Dahodwala - 2025 - Biotechnology Journal - Wiley Online Library

The following directive documents were developed at MCCC with support from a grant from the National Institute for Innovation in Manufacturing Biopharmaceuticals, NIIMBL.

1. ⇰ SOP Resuscitation and Culture of NISTCHO Cells
2. ⇰ SOP Cryopreservation of NISTCHO Cells
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4. Small Scale Culture
5. ⇰ SOP NISTCHO Shake Flask 30ml Batch Culture for Monoclonal Antibody Production
6. ⇰ SOP NISTCHO Batch Culture for Monoclonal Ab Production in T75 flask
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8. Upstream Processing
9. ⇰ NISTCHO Upstream Process Flow Diagram
10. ⇰ SOP Batch Culture of NISTCHO Cells for Production of cNISTmAb
11. ⇰ Batch Record Culture of NISTCHO cells for Production of cNISTmAb
12.  
13. Downstream Processing

14. ⇰ NISTCHO Downstream Process Flow Chart

• ⇰ SOP End of Run cNIST mAb Harvest, Centrifugation and TFF
• ⇰ Batch Record Downstream Processing of cNISTmAb
• ⇰ SOP Purification of cNISTmAb from Clarified Media by Protein A Chromatography on the AKTA System
• ⇰ SOP Purification of cNISTmAB from NISTCHO Clarified Media By Gravity Flow Protein A Chromatography

• ⇰ SOP Operation of Logos Biosystems Luna Fluorescence Cell Counter for Fluorescence Cell Counting
• ⇰ SOP Measurement of Glucose and Lactate Concentrations in Cell Culture Media using YSI 2500 Bioanalyzer
• ⇰ SOP Monoclonal Antibody Aggregate Testing by HPLC SEC Analysis

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1. Upstream Processing

2. ⇰  mAb Upstream Process Flow Chart

3. ⇰   Batch Record: Anti-IL-8 mAb Production from CHO DP-12 Cells
4. ⇰   Batch Culture of CHO DP12 Cells for production of anti-IL-8 Monoclonal Antibody SOP
5. ⇰   Bellco Spinner Flask (100mL) Cleaning and Autoclaving SOP
6. ⇰   Applikon ez-Control Bioreactor Controller Operation SOP
7.  
8. Upstream Quality Control
9. ⇰   Glucose Determination Assay Using Spectrophotometry SOP
10. ⇰   Lactate Determination Assay Using Spectrophotometry SOP
11. ⇰   Measurement of Glucose and Lactate Concentrations in Cell Culture Media using the YSI 2500 Bioanalyzer SOP
12. ⇰   Oakton PC 700 Bench Series pH/ Conductivity/°C/°F Meter SOP
13. ⇰   Operation of Life Technologies Countess II Cell Counter SOP
14. ⇰   Trypan Blue Assay for Cell Viability Determination SOP
15. ⇰   Quantitation of CHO DP-12 Derived Anti IL-8 Monoclonal Antibody by ELISA SOP
16.  
17. Downstream Processing
18. ⇰ FLOW CHART: mAb Downstream Process
19.  

20. Tangential Flow Filtration

21. ⇰ End of Run anti-IL8 mAb Process: Harvest, Centrifugation, TFF Concentration SOP
22. ⇰ BATCH RECORD: Downstream Processing of Anti-IL8 mAb
23.    
24. ÄKTA Purification System
25. ⇰ USER GUIDE: AKTA Pure 25 User Introduction
26. ⇰ Operation of ÄKTA pure Chromatography System SOP
27. ⇰ Isolation of mAb (anti-IL8)  from Conditioned Medium by Protein A
28. ⇰ Affinity Chromatography on the AKTA pure Chromatography System SOP
     
29.     
30. BioRad Biologics LP Purification System
31. ⇰ Biologic LP Chromatography System Operation SOP
32. ⇰ BATCH RECORD: Downstream Process Isolation of Anti-IL8 mAb from CHO Cells: Chromatography Operation
33. ⇰ Isolation of Anti-IL8 mAb by Protein A Affinity Chromatography SOP
34.  
35. Downstream Quality Control
36. ⇰ Fisher Scientific Vertical MiniGel System SOP
37. ⇰ SDS-PAGE SOP
38. ⇰ Quantitation of CHO DP-12 Derived Anti IL-8 Monoclonal Antibody by ELISA SOP
39. ⇰ Monoclonal Antibody Aggregate Testing by HPLC SEC Analysis SOP
40.  
41. Equipment Lists

42. ⇰   Equipment & Reagent List for Upstream Processing

43. ⇰   Equipment & Reagent List for Downstream Processing

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