High-Throughput Assessment of Structural Continuity in Biologics

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmaceutical Sciences

Background:

• Biologic drug products require consistent higher-order structure (tertiary and quaternary) for efficacy and safety.
• Assessing and ensuring this structural consistency is critical but challenging.
• Existing methods may not be sufficiently sensitive or high-throughput for comprehensive analysis.

Purpose of the Study:

• To demonstrate a high-throughput chemoprinting platform for assessing protein biologic structural consistency.
• To validate the platform's sensitivity in detecting subtle structural changes, including single-point mutations.
• To address the critical need for examining tertiary and quaternary protein structures in drug products.

Main Methods:

• Utilized a high-throughput chemoprinting platform employing small-molecule ligands as molecular probes.
• Developed libraries of probe molecules to generate holistic structural "fingerprints" for proteins.
• Applied the platform to enzymes and antibodies for proof-of-concept validation.

Main Results:

• Successfully confirmed consistency in the higher-order structure of protein biologics.
• Demonstrated high sensitivity capable of detecting single-point mutations.
• Identified minor conformational perturbations between protein mutants differing by a single amino acid residue.

Conclusions:

• The chemoprinting platform is effective for quality control of protein biologics.
• This method provides a sensitive and high-throughput approach to assess critical protein structural parameters.
• The platform enables the detection of subtle structural variations impacting biologic drug product consistency.