Use of peptide microarrays for fast and informative profiling of therapeutic antibody formulation conditions

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Abstract

Methods to optimize the solution behaviour of therapeutic proteins are frequently time consuming, provide limited information and often use milligram quantities of material. Here we present a simple, versatile method which provides valuable information to guide the identification and comparison of formulation conditions for, in principle, any biopharmaceutical drug. The subject protein is incubated with a designed synthetic peptide microarray; the extent of binding to each peptide is dependent on the solution conditions. The array is washed and adhesion of the subject protein detected using a secondary antibody. We exemplify the method using a well-characterized human single chain Fv and a selection of human monoclonal antibodies. Correlations of peptide adhesion profiles can be used to establish quantitative relationships between different solution conditions, allowing sub-grouping into dendrograms. Multidimensional reduction methods, such as t-distributed stochastic neighbor embedding, can be applied to compare how different monoclonals vary in their adhesion properties under different solution conditions. Finally, we screened peptide binding profiles using a selection of monoclonal antibodies for which a range of biophysical measurements were available under specified buffer conditions. We used a neural network method to train the data against aggregation temperature, kD, percentage recovery after incubation at 25oC and melting temperature. The results demonstrate that peptide binding profiles can indeed be effectively trained on these indicators of protein stability and self-association in solution. The method opens up multiple possibilities for the application of machine learning methods in therapeutic protein formulation.

Bibliographical metadata

Original languageEnglish
JournalMolecular Pharmaceutics
Publication statusPublished - 18 Oct 2021