Kim K J, Taylor D W, Evans C B, Asofsky R
J Immunol. 1980 Dec;125(6):2565-9.
A solid-phase radioimmunoassay (SPRIA) in microtiter wells was established for detecting antibodies against Plasmodium yoelii Ag. The SPRIA was found 1) to require as little as 5 micrograms of crude parasite Ag per well, 2) to be able to detect 0.5 ng of monoclonal Ab, and 3) to be 10(4) times more sensitive than the indirect fluorescent Ab staining technique. In a modification of the above assay using intact RBC as an Ag, hyperimmune serum showed significant binding to the surface of erythrocytes of mice infected with P. yoelii parasites but not to RBC of normal mice. Hybridomas were prepared by fusing infected mouse spleen cells with P3-NS1/I-Ag4-1 myeloma cells. Using the SPRIA, hybrids secreting Ab against P. yoelii 17XL Ag were detected. All the hybrids detected by SPRIA secreted Ab binding to free parasites but not to the surface membrane Ag on the infected RBC. The number of positive hybrids produced by fusing spleen cells from mice given a double infection (17XNL and 17 XL) was not greater than that produced by freezing spleen cells from mice given a single infection (17 XL).
建立了一种用于检测抗约氏疟原虫抗原抗体的微量滴定板固相放射免疫分析（SPRIA）。发现该SPRIA：1）每孔仅需5微克粗制寄生虫抗原；2）能够检测出0.5纳克单克隆抗体；3）灵敏度比间接荧光抗体染色技术高10⁴倍。在上述以完整红细胞作为抗原的分析方法的改进中，超免疫血清显示与感染约氏疟原虫的小鼠红细胞表面有显著结合，但与正常小鼠的红细胞无结合。通过将感染小鼠的脾细胞与P3-NS1/I-Ag4-1骨髓瘤细胞融合制备杂交瘤。使用SPRIA检测分泌抗约氏疟原虫17XL抗原抗体的杂交瘤。通过SPRIA检测到的所有杂交瘤均分泌与游离寄生虫结合的抗体，但不与感染红细胞表面膜抗原结合。用双重感染（17XNL和17XL）小鼠的脾细胞融合产生的阳性杂交瘤数量不大于用单一感染（17XL）小鼠的冷冻脾细胞产生的阳性杂交瘤数量。
