Engineered scPDL1-DM1 drug conjugate with improved in vitro analysis to target PD-L1 positive cancer cells and intracellular trafficking studies in cancer therapy

Abstract

Antibody-drug conjugates (ADC), precisely deliver a cytotoxic agent to antigen-expressing tumor cells by using specific binding strategies of antibodies. The ADC has shown the ability of potent bio-therapeutics development but indefinite stoichiometric linkage and full-length antibody penetration compromised the field of its advancement. Single chain variable fragments convention instead of the full-length antibody may overcome the challenge of rapid penetration and internalization. Programmed cell death ligand-1 interaction with PD-1 has recently revolutionized the field of immunotherapy. We systematically designed scPDL1-DM1 drug conjugate by linking scFv-PD-L1 proteins (scFv) with maytansinoids (DM1) cytotoxic agent through succinimidyl trans-4-maleimidylmethyl cyclohexane-1-carboxylate (SMCC) linker. Binding affinity was confirmed by immunocytochemistry, spectrophotometry and gel electrophoresis analysis. The scPDL1-DM1 showed specific binding with PD-L1 positive tumor cells and retained in vitro anti-cell proliferation activity. The intracellular trafficking of the drug was evaluated in A549 cancer cell lines, and maximum trafficking was observed after two hours of incubation. The generated drug can be utilized as a potent tool for site-specific conjugation, predicting specificity in vitro activities with extended range against PD-L1 positive cancer cells and can be utilized for further in vivo testing and clinical therapeutics development.

Bioengineering and computational analysis of programmed cell death ligand-1 monoclonal antibody

“…This method can be used to generate antibodies against tumor surface antigens and their ligands (34). We previously reported on the successful optimization of the extracellular domain of programmed cell death ligands-1, which was then processed to produce single chain variable fragments, scFv-PDL1 (35,36). The most common host cell machinery used in the manufacture of therapeutic monoclonal antibodies is CHO cells.…”

New Technologies Bloom Together for Bettering Cancer Drug Conjugates

“…It can identify desirable specificity and binding affinities. A recent study constructed an scFv antibody fragment-drug conjugate targeting PD-1 and conjugated with DM1 through a succinimidyl trans-4-maleimidylmethylcyclohexane-1-carboxylate linker, yielding potent antitumor activity and efficient intracellular trafficking results (Kalim et al, 2020). Single domain antibody fragments (single variable heavy chain or nanobodies) have simpler structures, improved solubility, high yield (50 mg/L) in Escherichia coli, and enhanced tissue penetration over full-sized antibodies.…”

Depleting Tumor Cells Expressing Immune Checkpoint Ligands—A New Approach to Combat Cancer

“…Several ADCs have gained regulatory approval and many others are in clinical development [ 182 , 183 ]. Anti-ICPML ADCs against several ICPMs (e.g., PD-L1, CD70, B7-H3, B7-H4, TIM-1) have been reported and some are in active clinical development while the development of some others has been discontinued because of marginal single-agent activity or unacceptable toxicity [ 131 , 139 , 145 , 146 , 147 , 148 , 149 , 150 , 151 , 152 , 153 , 154 , 155 , 156 ].…”