Development of an Influenza virus Protein Array Using Sortagging Technology

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• 作者：Antonia Sinisi ; Maximilian Wei-Lin Popp ; John M. Antos ; Werner Pansegrau ; Silvana Savino ; Mikkel Nissum ; Rino Rappuoli ; Hidde L. Ploegh ; Ludovico Buti
• 刊名：Bioconjugate Chemistry
• 出版年：2012
• 出版时间：June 20, 2012
• 年：2012
• 卷：23
• 期：6
• 页码：1119-1126
• 全文大小：362K
• 年卷期：v.23,no.6(June 20, 2012)
• ISSN：1520-4812

文摘

Protein array technology is an emerging tool that enables high-throughput screening of protein鈥損rotein or protein鈥搇ipid interactions and identification of immunodominant antigens during the course of a bacterial or viral infection. In this work, we developed an Influenza virus protein array using the sortase-mediated transpeptidation reaction known as 鈥淪ortagging鈥? LPETG-tagged Influenza virus proteins from bacterial and eukaryotic cellular extracts were immobilized at their carboxyl-termini onto a preactivated amine-glass slide coated with a Gly₃ linker. Immobilized proteins were revealed by specific antibodies, and the newly generated Sortag-protein chip can be used as a device for antigen and/or antibody screening. The specificity of the Sortase A (SrtA) reaction avoids purification steps in array building and allows immobilization of proteins in an oriented fashion. Previously, this versatile technology has been successfully employed for protein labeling and protein conjugation. Here, the tool is implemented to covalently link proteins of a viral genome onto a solid support. The system could readily be scaled up to proteins of larger genomes in order to develop protein arrays for high-throughput screening.