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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Appropriate coating methods and other conditions for enzyme-linked immunosorbent assay of smooth, rough, and neutral lipopolysaccharides of Pseudomonas aeruginosa.
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Appropriate coating methods and other conditions for enzyme-linked immunosorbent assay of smooth, rough, and neutral lipopolysaccharides of Pseudomonas aeruginosa.

机译：铜绿假单胞菌光滑，粗糙和中性脂多糖的酶联免疫吸附测定的适当包被方法和其他条件。

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摘要

Smooth, rough, and neutral forms of lipopolysaccharide (LPS) from Pseudomonas aeruginosa were used to assess the appropriate conditions for effective enzyme-linked immunosorbent assay (ELISA) of LPS. Each of these forms of well-defined LPS was tested for the efficiency of antigen coating by various methods as well as to identify an appropriate type of microtiter plate to use. For smooth LPS, the standard carbonate-bicarbonate buffer method was as efficient as the other sensitivity-enhancing plate-coating methods compared. The rough LPS, which has an overall hydrophobic characteristic, was shown to adhere effectively, regardless of the coating method used, to only one type of microtiter plate, CovaLink. This type of plate has secondary amine groups attached on its polystyrene surface by carbon chain spacers, which likely favors hydrophobic interactions between the rough LPS and the well surfaces. Dehydration methods were effective for coating microtiter plates with the neutral LPS examined, which is composed predominantly of a D-rhamnan. For the two dehydration procedures, LPS suspended in water or the organic solvent chloroform-ethanol was added directly to the wells, and the solvent was allowed to dehydrate or evaporate overnight. Precoating of plates with either polymyxin or poly-L-lysine did not give any major improvement in coating with the various forms of LPS. The possibility of using proteinase K- and sodium dodecyl sulfate-treated LPS preparations for ELISAs was also investigated. Smooth LPS prepared by this method was as effective in ELISA as LPS prepared by the hot water-phenol method, while the rough and neutral LPSs prepared this way were not satisfactory for ELISA.
机译：铜绿假单胞菌的脂多糖（LPS）的光滑，粗糙和中性形式用于评估LPS有效酶联免疫吸附测定（ELISA）的适当条件。通过各种方法测试了这些形式的定义明确的LPS的抗原包被效率，并确定了要使用的合适类型的微量滴定板。对于光滑的LPS，标准的碳酸盐-碳酸氢盐缓冲液方法与其他提高感光度的印版涂布方法一样有效。粗略的LPS具有总体疏水性，无论使用哪种涂布方法，都可以有效地粘附到一种类型的微量滴定板CovaLink上。这种类型的板具有通过碳链间隔基连接在其聚苯乙烯表面的仲胺基，这可能有利于粗糙LPS和孔表面之间的疏水相互作用。脱水方法对于用中性LPS（主要由D-rhamnan组成）检测的微量滴定板进行涂层是有效的。对于这两种脱水程序，将悬浮在水中或有机溶剂氯仿-乙醇中的LPS直接添加到孔中，并使溶剂脱水或蒸发过夜。用多粘菌素或聚-L-赖氨酸对板进行预涂层在用各种形式的LPS进行涂层中均未获得任何重大改进。还研究了将蛋白酶K和十二烷基硫酸钠处理的LPS制剂用于ELISA的可能性。用这种方法制得的光滑LPS在ELISA中与用热水酚法制得的​​LPS一样有效，而用这种方法制得的粗糙和中性LPS不能令人满意。

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