Lafky Jacqueline M, Baron Andre T, Maihle Nita J
Endocrine Research Unit, Mayo Clinic College of Medicine, Rochester, MN, USA.
Methods Mol Biol. 2006;327:39-47. doi: 10.1385/1-59745-012-X:39.
This chapter provides basic information for quantifying soluble epidermal growth factor receptor (sEGFR) isoforms in human sera using an acridinium-linked immunosorbent assay (ALISA) developed by Baron and coworkers. This ALISA has been shown to be specific for epidermal growth factor receptor (EGFR) and the isoforms of EGFR encoded by alternate transcripts (i.e., sEGFRs); it, therefore, does not detect other EGFR/ErbB receptor family members, i.e., ErbB2, ErbB3, or ErbB4. In addition, this ALISA recognizes EGFR and sEGFR isoforms that contain subdomains I-IV of the extracellular domain, but it does not recognize EGFR isoforms lacking subdomain IV. The ALISA described here also may be useful for determining EGFR and sEGFR concentrations in lysates of cultured cells, conditioned culture medium, or tissue/tumor specimens, as well as in other human body fluids such as serum, plasma, ascites, urine, saliva, and cystic fluids.
本章提供了使用由巴伦及其同事开发的吖啶鎓连接免疫吸附测定法（ALISA）对人血清中的可溶性表皮生长因子受体（sEGFR）亚型进行定量的基本信息。这种ALISA已被证明对表皮生长因子受体（EGFR）以及由可变转录本编码的EGFR亚型（即sEGFRs）具有特异性；因此，它不会检测其他EGFR/ErbB受体家族成员，即ErbB2、ErbB3或ErbB4。此外，这种ALISA识别包含细胞外结构域I-IV亚结构域的EGFR和sEGFR亚型，但不识别缺乏亚结构域IV的EGFR亚型。这里描述的ALISA也可用于测定培养细胞裂解物、条件培养基或组织/肿瘤标本以及其他人体体液（如血清、血浆、腹水、尿液、唾液和囊液）中的EGFR和sEGFR浓度。
