Biopesticides Regulation Services

1
ADC Downstream Purification Technology
Tel [phone] Email info_at_bocsci.com
2
Contents
www.adc.bocsci.com
Copyright 2022 BOC Sciences. All rights
reserved.
3
Basic Composition of ADC Drug

• Payload
• Sub-nanomolar toxicity
• Stable in target cells
• (Ideally) by-stander effect

• Monoclonal Antibody
• Target antigen selectively
• No immunogenicity
• Stable in plasma
• Internalizable

4
ADC Analysis Characterization
ADC Analysis and Characterization Methods
Chromatographic Analysis Of Drug Distribution
Size Exclusion Chromatography Analysis
Antibody Drug Conjugation Ratio (DAR)
Drug Distribution
For any given ADCs, the chemical property of
cytotoxins and linkers, as well as the choice of
attachment sites, will greatly affect the
physicochemical properties. Thus, different ADCs
may utilize different analytical assessment
methods according to their unique properties.
Charge Variant Separation
Free Small Molecule Analysis
Peptide Mapping
Biological Analysis
5
mAb
ADC Drug Production Process
Dilution/ Bulfer Exchange by TFF
Reaction mixture
mAb Modfication(ex. TCEP Reduction)
In the past 20 years, the research and
development of ADC drugs have become a hot spot
of competition among major pharmaceutical
companies. Currently, the FDA has approved more
than 10 ADC drugs, and more than 100 ADC drugs
are under clinical trials or pharmaceutical
research and development stages. Therefore, major
pharmaceutical companies must develop a stable
ADC platform-based production process.
mAb intermediate
Payood Conjugation
Crude ADC
Chromatography (SEC, HIC membrane etc.)
Purifcaton by TFF
Purified ADC
Purified ADC
Bufler Exchange/ Formulation by TFF
Butler Exchange/ Formulation by TFF
Final ADC composition
6
Tangential Flow Filtration (TFF)
Tangential flow filtration is a pressure-driven
membrane purification technique based on
molecular size. In tangential flow filtration,
the sample mixture is not forced through a single
passageway membrane as in direct flow filtration
but is recirculated tangentially across the
membrane surface through multiple cycles.
Materials Science in Semiconductor Processing.
2022, 145 106618.
7
Size Exclusion Chromatography (SEC)

• Buffer Exchange
• Aggregate Removal
• Non-protein Impurity Removal
• Small Molecular Impurity Removal

Journal of Pharmaceutical and Biomedical
Analysis. 2014, 101161-173.
8
Hydrophobic Interaction Chromatography (HIC)
ADC drugs are covalently coupled with hydrophobic
drug linkers, toxins, monoclonal antibodies, and
impurities that reveal different drug-to-antibody
ratios, and hydrophobic chromatography is used
according to the hydrophobic differences for
removal.
Journal of Pharmaceutical and Biomedical
Analysis. 2020, 186 113313.
9
Ion Exchange Chromatography (IEC)
Ion exchange chromatography involves protein
analysis, such as charge variant analysis and
glycoform analysis, to elucidate the structural
changes of proteins. Ion exchange chromatography
generally yields high recovery and removes
non-antibody-related compounds, including small
molecules and endotoxins. Therefore, ion exchange
chromatography is a promising method of ADC
purification.

• Small Molecule Impurities
• Host DNA Impurities
• Endotoxin Impurities
• Charge Variant Impurities

10
Hydroxyapatite Chromatography (HA)

• Hydroxyapatite chromatography is a useful method
  for monoclonal antibody purification.
• Hydroxyapatite chromatography utilizes calcium
  and phosphate minerals, which bind to antibodies
  through the interaction of calcium metal affinity
  and phosphate-based cation exchange.
• The use of hydroxyapatite chromatography can
  reduce antibody aggregates from 60 to 0.1.

The Crystalline Structure of HA
Ceramics, 2021, 4(4) 542-563.
11
Membrane Chromatography (MC)
J. Clin. Med. 2021, 10, 552.
12
Tel [phone] Email info_at_bocsci.com Addre
ss 45-16 Ramsey Road, Shirley, NY 11967, USA