J Chromatogr A. 2026 May 2;1780:467062. doi: 10.1016/j.chroma.2026.467062. Online ahead of print.

ABSTRACT

Biocatalysis, the use of enzymes to catalyze chemical transformations, is becoming an increasingly important focus in the manufacture of pharmaceutical drug substances. The use of biocatalysts lends marked improvements in process efficiency and sustainability, aligning with industry-wide initiatives for improving greenness. Despite this advantage, biocatalysts introduce additional challenges that must be addressed to ensure patient safety and product quality. Notably, control of biocatalyst residuals must be demonstrated through process optimization and nuanced analytical techniques. Analytical techniques have long been established for the quantification of protein, however in this context many fall short in terms of accuracy, user-friendliness, reproducibility, and susceptibility to matrix interferences. To address these shortcomings, an analytical approach using RPLC with UV absorbance detection to quantify biocatalyst residuals in the presence of drug substances and/or synthetic intermediates is presented. Preliminary data for the broad application to small molecule and oligonucleotide drug substance modalities were demonstrated. Further, the approach was validated as a method to quantify residual protein in a late-stage small molecule intermediate synthesized using a biocatalyst. The approach provides a user friendly, robust and broadly applicable solution that overcomes critical analytical limitations, thereby supporting the continued expansion of biocatalytic routes in pharmaceutical development and manufacturing.

PMID:42114345 | DOI:10.1016/j.chroma.2026.467062