A workflow to identify mutations of bulk recombinant proteins by using N-terminal labeling combined with the MELD method

机译：通过使用N-末端标记与MELD方法相结合鉴定块状重组蛋白突变的工作流程

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By using a N-terminal labeling step occurring at the protein level combined with the MultiEnzymatic Limited Digestion method (MELD) both frameshift and truncated mutations can be identified. A manual validation of the MSMS spectra quality is mandatory as no current FDR calculation for intra-peptide local mutation is available. Undesired co-produced forms were identified and relatively quantified across several experimental conditions. This analytical workflow can be used: 1. to optimize the production process; 2. to understand the biological events at the origin of the co-produced undesired forms.

机译：通过在蛋白质水平上发生的N-末端标记步骤，与多酶有限消化方法（MELD）相结合，可以识别架构和截断的突变。 MSMS光谱质量的手动验证是强制性的，因为没有目前肽内局部突变的FDR计算。在若干实验条件下鉴定并相对量化了不需要的共生形式。可以使用此分析工作流程：1。优化生产过程; 2.了解共同产生的不期望形式的起源的生物事件。

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《ASMS Conference on Mass Spectrometry and Allied Topics》|2016年|1CD|共1页
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Gabriel Mazzucchelli; Denis Morsa; Dominique Baiwir; Nicolas Smargiasso; Edwin De Pauw;
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2. Efficient sortase-mediated N-terminal labeling of TEV protease cleaved recombinant proteins [J] . Sarpong Kwabena, Bose Ron Analytical Biochemistry: An International Journal of Analytical and Preparative Methods . 2017,第期

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4. A workflow to identify mutations of bulk recombinant proteins by using N-terminal labeling combined with the MELD method [C] . Gabriel Mazzucchelli, Denis Morsa, Dominique Baiwir, ASMS Conference on Mass Spectrometry and Allied Topics . 2016

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5. Identification of Non-Random Somatic Mutation Clustering While Accounting for Protein Tertiary Structure: Extensions, Novel Methodologies and Applications to Identifying Oncogenic Driver Mutations. [D] . Ryslik, Gregory A. 2014

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6. Efficient Sortase-mediated N-terminal Labeling of TEV Protease Cleaved Recombinant Proteins [O] . Kwabena Sarpong, Ron Bose -1

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A workflow to identify mutations of bulk recombinant proteins by using N-terminal labeling combined with the MELD method

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