De-Simone Salvatore G, Napoleão-Pêgo Paloma, De-Simone Thatiane S
FIOCRUZ, Center of Technological Development in Health (CDTS) / National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), FIOCRUZ, Rio de Janeiro, Brazil.
Department of Cellular andMolecularBiology, Federal FluminenseUniversity,Biology Institute, Niterói, Rio de Janeiro, Brazil.
Methods Mol Biol. 2016;1352:263-77. doi: 10.1007/978-1-4939-3037-1_20.
Peptide microarrays have become increasingly more affordable in recent years with the SPOT technique being one of the most frequently used methods for synthesis and screening of peptides in arrays. Here, a protocol is presented for the identification of the amino acid sites involved in the conversion of human IgG to IgE response during the passive administration of therapeutic, anti-snake venom sera. Similarly, the minimal region of both the IgG and IgE binding epitopes, important for its interaction with ligand, were identified. As the ratio of concentrations for IgG to IgE in human serum is 1:10,000, also presented is a reproductive protocol of chemiluminescence-scanning for the detection of both immunoglobulins.
近年来，肽微阵列的价格越来越亲民，其中SPOT技术是阵列中肽合成和筛选最常用的方法之一。本文介绍了一种用于鉴定在被动给予治疗性抗蛇毒血清过程中参与人IgG向IgE反应转化的氨基酸位点的方案。同样，还确定了IgG和IgE结合表位中对其与配体相互作用很重要的最小区域。由于人血清中IgG与IgE的浓度比为1:10000，本文还介绍了一种用于检测这两种免疫球蛋白的化学发光扫描重现性方案。
