Dual Site-Specific Antibody Conjugates for Sequential and Orthogonal Cargo Release

Area of Science:

• Bioconjugation Chemistry
• Antibody Engineering
• Drug Delivery Systems

Background:

• Antibody-drug conjugates (ADCs) offer targeted therapy by combining antibody specificity with potent drug payloads.
• Drug resistance necessitates advanced delivery systems for multidrug cocktails.
• Site-specific conjugation enhances ADC homogeneity and therapeutic efficacy.

Purpose of the Study:

• To engineer a novel dual-labeled antibody bioconjugate platform for controlled, sequential drug release.
• To demonstrate orthogonal cargo release mechanisms for enhanced therapeutic strategies.
• To establish temporal control over drug delivery from a single antibody conjugate.

Main Methods:

• Engineered lipoate-acid ligase A (LplA) acceptor peptide (LAP) tags into trastuzumab.
• Utilized chemoenzymatic ligation with microbial transglutaminase (mTG) and LplA for site-specific conjugation.
• Achieved orthogonal cargo release via matrix metalloproteinase-2 and cathepsin-B cleavage.

Main Results:

• Successfully generated dual-labeled antibody bioconjugates with engineered LAP tags and Q295 residue.
• Demonstrated orthogonal and sequential release of distinct drug payloads.
• Established temporal control over drug release from the engineered antibody platform.

Conclusions:

• This work presents the first demonstration of temporal control in dual-labeled antibody conjugates.
• The developed platform enables sequential drug release and cooperative drug combinations.
• This technology holds promise for advanced targeted therapies against drug-resistant cancers and infections.