Simultaneous LC-MS/MS Quantification Method for Cassette-Dosing Pharmacokinetics Study of Monoclonal Antibodies Using...

Area of Science:

• Biochemistry
• Pharmacology
• Immunology

Background:

• Antibody therapeutics, particularly IgG1, are crucial in medicine.
• Evaluating pharmacokinetics (PK) of multiple antibodies simultaneously is challenging.
• Existing methods can be time-consuming and costly, impacting drug development timelines.

Purpose of the Study:

• To develop a novel cassette-dosing PK evaluation method for IgG1 antibodies.
• To enable simultaneous measurement of multiple engineered antibodies recognizing the same antigen.
• To assess the impact of antibody engineering on PK profiles and validate the method's efficiency and reliability.

Main Methods:

• Antibody engineering: alanine mutations/insertions in the CH1 domain (GPSVFPLAPSSK sequence) of IgG1 antibodies.
• Multiplex liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous quantification of engineered antibodies.
• Cassette-dosing PK studies in mice followed by comparison with single-dosing PK data.

Main Results:

• Validated assay performance with accuracy (70.3%-128.4%) and precision (%CV ≤ 28.7%).
• Engineered antibodies exhibited comparable PK profiles (t1/2, C0, AUC(last), Vss) to unmodified antibodies in mice.
• Cassette-dosing PK results were consistent with single-dosing studies, indicating no drug-drug interactions.

Conclusions:

• The novel cassette-dosing PK method efficiently evaluates multiple IgG1 antibodies.
• Antibody engineering modifications did not substantially impact PK profiles.
• This approach accelerates lead candidate selection, predicts human PK, reduces animal use (3Rs), and lowers drug development costs.