HEBREW UNIV JERUSALEM HOSP,KAPLAN & HADASSAH,REHOVOT,ISRAEL. HEBREW UNIV JERUSALEM HOSP,KORET SCH VET MED,LAB TERATOL & EXPT ONCOL,REHOVOT,ISRAEL. HEBREW UNIV JERUSALEM HOSP,FAC AGR,DEPT BIOCHEM,REHOVOT,ISRAEL.
Int J Oncol. 1996 Jul;9(1):153-7.
Gel fiberglass (GFG), a new affinity biosensor, was used to isolate human p53 antigen with rabbit anti-rat p53 IgG. The biosensor was prepared as a membrane from glass fibers covered with oxysilanes. A thin layer of protein, trapped in gel glass during its preparation, is deposited on the surface of a lattice of glass fibers. In such conditions, a maximum number of protein molecules may contact external agents percolated through a membrane. The membranes demonstrate high stability and can be stored in dry conditions or several months at room temperature. Columns for affinity chromatography were prepared from the GFG membranes and were used to isolate various proteins, including the tumor-associated antigens (TAA). The capacity of such columns was calculated as the amount mg of protein isolated from 1 ml of TAA-containing serum. In colon cancer patients, up to 5-6 mg TAA were extracted from 1 mi of sera. Two main components of cytoplasmic TAA isolated in our experiments were p64 and p53 proteins. Their concentration was determined by HPLC. The p53 protein has been isolated from the serum of cancer patients in the highest concentration yet reported, up to 3-4 mg/ml. In our previous studies, isolation of p53 protein was based on its affinity reaction with anti-p53 IgG generated against antigens of the same species. Herein, we report for the first time the capability to isolate human p53 antigen using GFG columns with entrapped anti-rat p53 IgG. Blood levels of p53 antigen isolated were very similar in both experiments. This has both theoretical and practical significance, demonstrating that the GFG membranes have great potential for isolating macromolecules utilizing various ligands. The finding facilitates an easy and highly effective method to isolate antigens from different organs, both animal and human, which can be used for important goals including diagnosis, therapy and generation of specific antibodies.
凝胶玻璃纤维（GFG）是一种新型亲和生物传感器，用于使用兔抗鼠 p53 IgG 分离人 p53 抗原。该生物传感器由涂有氧化硅烷的玻璃纤维制成的膜制备而成。在制备过程中，蛋白质的薄层被困在凝胶玻璃中，并沉积在玻璃纤维晶格的表面上。在这种情况下，可能有最大数量的蛋白质分子与渗透过膜的外部试剂接触。这些膜具有很高的稳定性，可以在干燥条件下或在室温下储存数月。从 GFG 膜制备亲和色谱柱，并用于分离各种蛋白质，包括肿瘤相关抗原（TAA）。这种柱子的容量计算为从 1ml 含有 TAA 的血清中分离出的蛋白质的量（mg）。在结肠癌患者中，从 1ml 血清中提取高达 5-6mg 的 TAA。在我们的实验中分离的细胞质 TAA 的两个主要成分是 p64 和 p53 蛋白。它们的浓度通过 HPLC 确定。p53 蛋白已从癌症患者的血清中以迄今为止报道的最高浓度分离出来，高达 3-4mg/ml。在我们之前的研究中，p53 蛋白的分离是基于其与针对同种抗原产生的抗 p53 IgG 的亲和反应。在此，我们首次报道了使用包埋抗鼠 p53 IgG 的 GFG 柱分离人 p53 抗原的能力。两种实验中分离的 p53 抗原的血液水平非常相似。这具有理论和实际意义，表明 GFG 膜具有利用各种配体分离大分子的巨大潜力。这一发现为从不同器官（包括动物和人类）中分离抗原提供了一种简单而高效的方法，可用于包括诊断、治疗和产生特异性抗体在内的重要目标。
