Epitope screening of the PCV2 Cap protein by use of a random peptide-displayed library and polyclonal antibody

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• 作者：Meng Ge ; Ai Yan ; Wei Luo ; Yun-Fei Hu ; Run-Cheng Li ; Da-Liang Jiang ; Xing-Long Yu
• 关键词：Porcine circovirus 2 ; Capsid protein ; Phage display ; Epitope mapping ; Polyclonal antibodies
• 刊名：Virus Research
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：177
• 期：1
• 页码：103-107
• 全文大小：747 K

文摘

Capsid protein (Cap), the only structural protein of porcine circovirus 2 (PCV2), is involved in the host protective response and is a target for vaccine development. To find a rapid and easy way to fully map the antigenic epitopes of Cap, purified Cap-specific polyclonal antibodies were used to screen a random heptapeptide phage display library. After three rounds of screening, twenty phage clones that had binding activity to Cap-specific antibodies (tested by phage ELISA) were sequenced. When the inserted amino acid sequences were aligned with the Cap protein sequence, eight core regions in Cap (⁵⁰SRTFGYT⁵⁶, ⁶²VRTPSW⁶⁷, ⁶⁸AVDMMR⁷³, ⁷⁹FLPPGG⁸⁴, ⁸⁶SNPRSVPF⁹³, ¹⁰²KVEFWP¹⁰⁷, ¹¹⁹GSSXXXLDDN¹²⁸ and ²²⁹PPLNP²³³) were identified, three of which (⁵⁰SRTFGYT⁵⁶, ⁸⁶SNPRSVPF⁹³ and ¹⁰²KVEFWP¹⁰⁷) for the first time. Nine phagetopes representing the eight regions were chosen to immunize Kunming mice. All except minotopes ⁵⁰SRTFGYT⁵⁶ and ²²⁹PPLNP²³³ induced antibodies against PCV2 when injected into Kunming mice.