Grubb A, Löfberg H, Thysell H, Ljunggren L, Olsson T, Skinner M, Shirahama T, Cohen A S
Department of Clinical Chemistry, University of Lund, Malmö General Hospital, Sweden.
Scand J Clin Lab Invest. 1987 Oct;47(6):619-26.
The hydrophilic nonapeptide Ser-Asp-Ala-Arg-Glu-Asn-Ile-Gln-Arg, identical with residues 59-67 of human amyloid protein A (AA) and serum amyloid protein A (SAA), was covalently bound via its carboxyl-terminal end to the carrier-protein keyhole limpet haemocyanin. The complex was injected subcutaneously into ten rabbits. All rabbits produced antisera which, unabsorbed, were specific for AA and SAA. The antisera and their isolated peptide specific antibodies were performance-tested and found to be excellent for demonstration of AA and SAA in immunoblotting and immunohistochemical techniques but unsuitable for immunoprecipitation. Since it is difficult to produce AA- and SAA-specific antisera by procedures earlier described and commercial supplies of good such reagents are unavailable, the easy production of sequence-specific such antisera will facilitate more extended studies of the corresponding antigens for diagnostic and scientific purposes.
亲水性九肽丝氨酸-天冬氨酸-丙氨酸-精氨酸-谷氨酸-天冬酰胺-异亮氨酸-谷氨酰胺-精氨酸,与人淀粉样蛋白A(AA)和血清淀粉样蛋白A(SAA)的59 - 67位残基相同,通过其羧基末端与载体蛋白钥孔血蓝蛋白共价结合。该复合物皮下注射到10只兔子体内。所有兔子都产生了抗血清,未吸收的抗血清对AA和SAA具有特异性。对这些抗血清及其分离的肽特异性抗体进行了性能测试,发现它们在免疫印迹和免疫组织化学技术中用于检测AA和SAA非常出色,但不适用于免疫沉淀。由于通过先前描述的方法难以产生AA和SAA特异性抗血清,且市场上没有此类优质试剂的商业供应,因此这种序列特异性抗血清的简便生产将有助于为诊断和科学目的对相应抗原进行更广泛的研究。