Combining Machine Learning and Backgrounded Membrane Imaging: A Case Study in Comparing and Classifying Different Types of Biopharmaceutically Relevant Particles.

This study investigates how backgrounded membrane imaging (BMI) can be used in combination with convolutional neural networks (CNNs) in order to quantitatively and qualitatively study subvisible particles in both protein biopharmaceuticals and samples containing synthetic model particles. BMI requires low sample volumes and avoids many technical complications associated with imaging particles in solution, e.g., air bubble interference, low refractive index contrast between solution and particles of interest, etc. Hence, BMI is an attractive technique for characterizing particles at various stages of drug product development. However, to date, the morphological information encoded in brightfield BMI images has scarcely been utilized. Here we show that CNN based methods can be useful in extracting morphological information from (label-free) brightfield BMI particle images. Images of particles from biopharmaceutical products and from laboratory prepared samples were analyzed with two types of CNN based approaches: traditional supervised classifiers and a recently proposed fingerprinting analysis method. We demonstrate that the CNN based methods are able to efficiently leverage BMI data to distinguish between particles comprised of different proteins, various fatty acids (representing polysorbate degradation related particles), and protein surrogates (NIST ETFE reference material) only based on BMI images. The utility of using the fingerprinting method for comparing morphological differences and similarities of particles formed in distinct drug products and/or laboratory prepared samples is further demonstrated and discussed through three case studies.