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ADCs Conjugation

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ADCs Conjugation

Conventional methods:

Lysine site conjugation
Reduced cysteine site conjugation method

Site specific methods:

Engineered cysteine residue
Disulphide re-bridging
Unnatural amino acid
Enzyme assisted ligation
Glycan remodelling
P-click technology

Antibodies conjugation and characterization work handled- Trastuzumab (Her-2), Cetuximab (EGFR), Sacituzumab (TROP-2), Enfortumab (Nectin-5)

ADC services include antibody expression, conjugation, linker synthesis, characterization and efficacy studies

ADCs characterization

ADC-antigen affinity-

Cellular affinity through FACS
Cell-free affinity through SPR (antigen, FcγRs and FcRn) and ELISA.
Affinity comparison between conjugated and un-conjugated antibody

Antigen expression validation- Screening of different cell lines for antigen expression through FACS

Cell viability-

2D and 3D format
Competition cell viability (for target mediated potency confirmation, in presence of unconjugated antibody)
Cytotoxicity through LDH
Apoptosis assessment through Annexin V-FITC and propidium iodide staining.
Off-target cytotoxicity target-negative cell lines

ADC internalization- pH sensitive dye conjugation and detection through FACS/Fluorescent plate reader

In vivo anti-tumor efficacy of ADCs

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