人类黑色素瘤相关抗原：一种检测特异性抗体的固相测定法。

Human melanoma associated antigens: a solid-phase assay for detection of specific antibody.

作者信息

Morgan A C, Galloway D R, Wilson B S, Reisfeld R A

出版信息

J Immunol Methods. 1980;39(3):233-46. doi: 10.1016/0022-1759(80)90058-7.

Abstract

A solid-phase radioimmunometric binding assay is described utilizing 125I-labeled protein A for the detection of antibody to human melanoma associated antigens. The novel aspect of this assay is the use of chemically defined spent culture medium of melanoma cells at target antigens previously depleted of fibronectin by affinity chromatography. This makes it possible to screen for antibody in unabsorbed antiserum. Sensitivity, reproducibility and ease of performance of the assay are optimized by conjugating target antigens to a background of bovine serum albumin dried onto polyvinyl 96-well microtiter plates and cross-linked with glutaraldehyde. The use of an immobilized soluble antigen target derived from a large pool of spent culture medium facilitates direct interassay comparisons and permits extensive absorption analysis of antisera. The assay has considerable potential in screening for alloantibody and both poly- and monoclonal xeonoantibody to human melanoma associated antigens.

摘要

描述了一种固相放射免疫测定结合试验，利用125I标记的蛋白A检测人黑色素瘤相关抗原的抗体。该试验的新颖之处在于使用经化学定义的黑色素瘤细胞用过的培养基作为靶抗原，该靶抗原先前已通过亲和层析去除了纤连蛋白。这使得在未吸收的抗血清中筛选抗体成为可能。通过将靶抗原与干燥在聚乙烯96孔微量滴定板上并用戊二醛交联的牛血清白蛋白背景结合，优化了试验的灵敏度、重现性和操作简便性。使用源自大量用过的培养基的固定化可溶性抗原靶标有助于直接进行试验间比较，并允许对抗血清进行广泛的吸收分析。该试验在筛选针对人黑色素瘤相关抗原的同种抗体以及多克隆和单克隆异种抗体方面具有相当大的潜力。