EVALUATION OF THE DOMINANT PROTEIN FROM WHOLE-CELL HEATING METHOD USING MALDI-TOF/TOF MASS SPECTROMETRY

S. Ningrum, I. Wibawan
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Abstract

Immunoglobulin G (IgG) are the essential tools in diagnostic kits. The whole-cell is still considered to be used for animal‘s immunization to produce IgG. The goal of this study was to determine dominant protein on the Shiga toxin-producing Escherichia coli (STEC) O157:H7-induced typical antibody response also determined by MALDI-TOF/TOF mass spectrometry. The heating method was conducted at 71 °C and 83 °C for antigen preparation. The proteins were characterized using SDS-PAGE and evaluated by MALDI-TOF/TOF mass spectrometry. The present study reported that the dominant polyclonal antibody inducing protein was ompC E. coli. The evaluation of mass spectra by MALDI-TOF/TOF might help to identify the dominant polyclonal antibody inducing protein in future preparation of diagnostic kits.
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利用maldi-tof / tof质谱法对全细胞加热法的优势蛋白进行评价
免疫球蛋白G (IgG)是诊断试剂盒中必不可少的工具。全细胞法仍被认为可用于动物免疫以产生IgG。本研究的目的是确定产志贺毒素大肠杆菌(STEC) O157: h7诱导的典型抗体反应的优势蛋白,并通过MALDI-TOF/TOF质谱法测定。加热方法分别在71°C和83°C进行抗原制备。采用SDS-PAGE和MALDI-TOF/TOF质谱法对蛋白进行鉴定。本研究报道了主要的多克隆抗体诱导蛋白为ompC大肠杆菌。MALDI-TOF/TOF质谱评价有助于鉴定多克隆抗体诱导蛋白的优势蛋白,为今后制备诊断试剂盒提供参考。
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