Author(s): Dick LW Jr, Kakaley JA, Mahon D, Qiu D, Cheng KC, Dick LW Jr, Kakaley JA, Mahon D, Qiu D, Cheng KC
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Abstract Hydrolysates play an important role in modern biological production. These mixtures are mostly undefined and contain a mixture of proteins, peptides, and amino acids along with other non-amino acid-based components. Recently, there has been an interest in defining and sequencing proteins and peptides in these hydrolysates to subsequently develop an assay to ensure removal during product purification. This work investigates an ultrafiltrate of yeastolate to determine whether any protein is present. Size exclusion chromatography indicated a possible high molecular weight component (>10 kDa). This suspected high molecular weight fraction was collected and investigated. It was determined that this fraction consists of nucleic acids; and no protein was detected using sensitive modern techniques including HPLC, mass spectrometry, and SDS-PAGE. Next, five unique, yeast-specific peptides were identified, sequenced, and confirmed. Finally, an impurity assay for any residual yeast specific peptides was developed and the analytical metrics were determined including accuracy, precision, linearity, range, and limits of detection and quantitation. (c) 2009 American Institute of Chemical Engineers Biotechnol.
This article was published in Biotechnol Prog
and referenced in Journal of Bioprocessing & Biotechniques