The biopharmaceutical industry continues to develop protein-based biotherapeutics in increasing numbers. Due to their complexity and biotechnological production, there are many attributes that need to be analyzed to guarantee their safety and efficacy. Peptide mapping is used to measure several critical quality attributes (CQA) required for the characterization of any biotherapeutic protein. The analysis is used to confirm that the correct sequence has been expressed for the protein and to check for post-translational and chemical modifications.

The Thermo Scientific Vanquish H system with fluorescence detection is applied for the glycan profile comparison of a biosimilar candidate produced ‘in house’ using Chinese hamster ovary (CHO) cell line using HILIC-based chromatographic separation on Accucore 150-Amide-HILIC column. Developed methodology offers an excellent analytical platform for glycan profile comparison of multiples lots of the biosimilar product.

The Thermo Scientific Vanquish Flex UHPLC System was used in combination with the MAbPac RP column for the reversed-phase analysis of intact bevacizumab. Column thermostatting strongly influences peak shape and recovery. For runs with column temperature set between 60 and 90 °C, peak width at half height is below 3 seconds for a 30 minutes long gradient. The largest peak area is obtained at 70 °C, and decreases steeply at higher and lower column temperatures. Carry-over decreases at higher temperature. At 70 °C the carry-over is estimated at 0.37%.

Fermentation broths are complex mixtures of nutrients, waste products, cells and cell debris, and desired products. Fermentation monitoring is important for many industries because carbon sources and metabolic by-products can impact the yield of the desired products. This application note describes the use of two different anion-exchange columns, IonPac AS11 and IonPac AS11-HC, with suppressed conductivity detection, to analyze common organic and inorganic anions in yeast and bacterial fermentation broths.

Peptide mapping is one of the major analyses in biotherapeutics characterization. Here we show peptide mapping with the Thermo Scientific Vanquish Flex UHPLC system. An efficient and reliable separation of peptides obtained from a bovine serum albumin (BSA) digest is done on an Thermos Scientific Acclaim RSLC 120, C18, 2.2 µm Analytical column. Excellent retention time and area precisions allow reliable qualitative and quantitative data interpretation.