Ball J M, Henry N L, Montelaro R C, Newman M J
Division of Molecular Virology, Baylor College of Medicine, Houston, TX.
J Immunol Methods. 1994 May 2;171(1):37-44. doi: 10.1016/0022-1759(94)90226-7.
A simple, versatile and very inexpensive procedure for cross-linking synthetic peptides to the polystyrene surfaces of micro-well assay plates for use in ELISA was developed. The method is based on the use of poly-L-lysine (PLL) as the anchor protein for synthetic peptides which were then easily and covalently linked to the PLL using glutaraldehyde. The synthetic peptides used for the study were based on the amino acid sequence of the equine infectious anemia virus (EIAV) envelope sequence and evaluated as antigens in an ELISA designed to detect antibodies in serum of EIAV-infected horses and ponies. The ELISA using cross-linked peptides proved to be significantly more sensitive when compared to assays where passively coated peptides were used. In one instance, a peptide was identified that was not recognized by any of our antisera and appeared not to bind to the assay plates. However, once this peptide was cross-linked to the assay plate it proved to be very useful for detecting EIAV-specific antibodies. This cross-linking approach functioned equally well with peptides of various charges and sizes and did not appear to alter epitopes contained in the peptides.
开发了一种简单、通用且非常廉价的方法，用于将合成肽交联到微孔检测板的聚苯乙烯表面，以用于酶联免疫吸附测定（ELISA）。该方法基于使用聚-L-赖氨酸（PLL）作为合成肽的锚定蛋白，然后使用戊二醛将合成肽轻松且共价地连接到PLL上。用于该研究的合成肽基于马传染性贫血病毒（EIAV）包膜序列的氨基酸序列，并在旨在检测EIAV感染的马和小马血清中抗体的ELISA中作为抗原进行评估。与使用被动包被肽的检测方法相比，使用交联肽的ELISA被证明具有更高的灵敏度。在一个实例中，鉴定出一种肽，我们所有的抗血清均未识别该肽，并且它似乎不与检测板结合。然而，一旦将该肽交联到检测板上，它就被证明对检测EIAV特异性抗体非常有用。这种交联方法对各种电荷和大小的肽均同样有效，并且似乎不会改变肽中所含的表位。
