Cy5 labeled antimicrobial peptides for enhanced detection of Escherichia coli O157:H7
Cy5 labeled antimicrobial peptides for enhanced detection of Escherichia coli O157:H7
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DOI：
10.1016/j.bios.2008.02.005
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发表时间：
2008-06-15
影响因子：
12.6
通讯作者：
Senecal, Andre
中科院分区：
文献类型：
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作者：
Arcidiacono, Steven;Pivarnik, Philip;Senecal, Andre
在检测大肠杆菌O157：H7的夹心试验中，荧光标记的抗菌肽作为标记抗体的潜在替代品进行了评估。作为其作用模式的一部分，抗菌肽自然地与细菌细胞壁的脂多糖成分结合。由于其相对于抗体的尺寸较小，因此能够以更大的密度结合到细胞表面，从而增加了光信号并提高了灵敏度。这种方法结合了捕获抗体的特异性和使用标记的多肽作为检测分子所提供的更高的灵敏度。通过马来酰亚胺连接物化学方法，用荧光染料Cy5标记抗菌肽Cecropin PI、SMAP29和PgQ。用全细胞溶液结合试验初步筛选发现，相对于Cy5标记的抗E-E，Cy5抗菌肽PI增强了对大肠杆菌O157：H7的检测。ColiO157：H7抗体10倍。抗体和多肽的检测灵敏度也与免疫磁珠生物传感器样机进行了比较。使用Cy5天蚕素PI检测的灵敏度提高了10倍。多肽抗菌活性与大肠杆菌O157：H7检测的相关性表明，活性不能预测荧光检测的敏感性。爱思唯尔出版公司(Elsevier B.V.)
Fluorescently labeled antimicrobial peptides were evaluated as a potential replacement of labeled antibodies in a sandwich assay for the detection of Escherichia coli O157:H7. Antimicrobial peptides naturally bind to the lipopolysaccharide component of bacterial cell walls as part of their mode of action. Because of their small size relative to antibodies peptides can bind to cell surfaces with greater density, thereby increasing the optical signal and improving sensitivity. This method combines the specificity of a capture antibody with the increased sensitivity provided by using a labeled peptide as a detection molecule. The antimicrobial peptides cecropin PI, SMAP29, and PGQ were labeled with the fluorescent dye Cy5 via maleimide linker chemistry. Preliminary screening using a whole-cell solution binding assay revealed that Cy5 cecropin PI enhanced the detection of E. coli O157:H7 relative to a Cy5 labeled anti-E. coli O157:H7 antibody 10-fold. Detection sensitivity of antibody and peptide were also compared with a prototype immuno-magnetic bead biosensor. Detection using Cy5 cecropin PI resulted in a 10-fold improvement in sensitivity. Correlation of peptide antimicrobial activity with detection of E. coli O157:H7 indicated that activity was not predictive of the sensitivity of the fluorescent assay. Published by Elsevier B.V.