Competition-based cellular peptide binding assays for 13 prevalent HLA class I Alleles using fluorescein-labeled synthetic peptides
Competition-based cellular peptide binding assays for 13 prevalent HLA class I Alleles using fluorescein-labeled synthetic peptides
复制标题
DOI：
10.1016/s0198-8859(02)00787-5
复制
发表时间：
2003-02-01
期刊：
影响因子：
2.7
通讯作者：
Drijfhout, JW
中科院分区：
文献类型：
--
作者：
Kessler, JH;Mommaas, B;Drijfhout, JW
我们报道了13个流行的人类白细胞抗原(HLA1)类等位基因的基于竞争的多肽结合分析的发展、验证和应用。这些分析是基于携带特定等位基因的活细胞上与人类白细胞抗原分子结合的多肽。目的测试肽与荧光素标记的HLA1类结合肽之间的结合竞争被用于读出。使用细胞膜结合的人类白细胞抗原I类分子避免了以可溶性形式对这些分子进行费力的生化纯化的需要。此前，我们已经将这一原则应用于人类白细胞抗原A2和人类白细胞抗原A3。我们现在描述的是人类白细胞抗原A1、A11、A24、A68、B7、B8、B14、B35、B60、B61和B62的检测方法。加上人类白细胞抗原A2和人类白细胞抗原A3，这些等位基因覆盖了超过95%的高加索人群。每项检测都确定了几个等位基因特异的参数。利用这些分析方法，我们从HIVpol、P53、PRAME和次要组织相容性抗原HA-1中鉴定了新的人类白细胞抗原I类高亲和力结合肽。因此，这些简便而准确的多肽结合分析方法将有助于识别存在于不同的HLAI类分子阵列上的可能的细胞毒性T淋巴细胞表位。(C)美国组织相容性和免疫遗传学学会，2003年。爱思唯尔科学公司出版。
We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class 1 alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class 1 binding peptide is used as read out. The use of cell membrane-bound HLA class I molecules circumvents the need for laborious biochemical purification of these molecules in soluble form. Previously, we have applied this principle for HLA-A2 and HLA-A3. We now describe the assays for HLA-A1, HLA-A11, HLA-A24, HLA-A68, HLA-B7, HLA-B8, HLA-B14, HLA-B35, HLA-B60, HLA-B61, and HLA-B62. Together with HLA-A2 and HLA-A3, these alleles cover more than 95% of the Caucasian population. Several allele-specific parameters were determined for each assay. Using these assays, we identified novel HLA class I high-affinity binding peptides from HIVpol, p53, PRAME, and minor histocompatibility antigen HA-1. Thus these convenient and accurate peptide-binding assays will be useful for the identification of putative cytotoxic T lymphocyte epitopes presented on a diverse array of HLA class I molecules. (C) American Society for Histocompatibility and Immunogenetics, 2003. Published by Elsevier Science Inc.