巨噬細胞株 P388D1 可分泌抑制素抑制漿細胞瘤細胞株 MOPC-315 的生長
並誘發細胞自戕現象。 本實驗製備抗漿細胞瘤抑制素單株抗體並分析其特
性，隨後進一步以此單株抗體純化漿細胞瘤抑制素。P388D1 培養基上清液
，經 DEAE-Sephacel 離子交換色層分析，得到二群具有抑制活性的蛋白，
分別稱之為 Peak 1，Peak 2。隨後以 Peak 1 及 Peak 2 免疫 Balb/c 老
鼠，經脾臟細胞與骨髓瘤細胞株融合產生融合瘤之後，以中和 PIF 抑制活
性的生物測試及酵素免疫分析法進行篩選， 最後篩選出 38 株製造抗漿細
胞瘤抑制素單株抗體的融合瘤。其中以 Peak 1 和 Peak 2 一起免疫，獲
得 2 株單株抗體；以 Peak 1 免疫，獲得 28 株單株抗體；以 Peak 2 免
疫，獲得 8 株單株抗體。 CA5-1.23 ( gamma-1, lambda ; 效價為 3.27x
10000﹚，CB7-1.1﹙ gamma-1,lambda;效價為 5.24x100000﹚，CB7-1.5﹙
gamma-1,lambda;效價為 1.31x100000﹚，CB7-1.19﹙ gamma-1,lambda;效
價為 1.31x100000﹚，EE6-2.2﹙gamma-1,kappa;效價為 5.24x100000﹚等
五株抗體，進一步作西方點墨法分析，結果皆辨識 Peak 1 中之一分子量
約 60KDa 的蛋白；CB2-2.18﹙ mu,kappa;效價為 1.31x100000﹚，作西方
點墨法分析，結果辨識 Peak 1 中分子量約 60KDa、51KDa、50KDa、47KDa
等四種蛋白。 在 P388D1 和 MOPC-315 共同培養情況下，加入此六株單株
抗體，可中和 P388D1 對 MOPC-315 產生之抑制作用。 利用 CA5-1.23 單
株抗體製備親和性色層分析管，進一步純化漿細胞瘤抑制素， 純化倍率為
1045 倍，高純度之漿細胞瘤抑制素只需 0.625ng，便能抑制 MOPC-315 的
生長。

P388D1, macrophage-like cells secreted a plasmacytoma inhibitory
factor (PIF) that suppressed the growth of MOPC-315 and induced
apoptosis. The specific aim of this study was to prepared a panel
of monoclonal antibodies that neutralized the activity of PIF.
P388D1 culture supernatants were analyzed by a DEAE-Sephacel
chromatography, and the plasmacytoma inhibitory activity was
localized on two protein peaks designated as Peak 1 and Peak 2.
Both Peak 1 and Peak 2 were used to immunize Balb/c mices,
together and separately. The hybirdoma were prepared by fusing
spleen cells with myeloma and were screened using both
neutralization assay and ELISA. Among the 38 positive lines, 2
lines were obtained from Peak 1 & Peak 2 immunization, 28 lines
were obtained from Peak 1 immunization, 8 lines were obtained
from Peak 2 immunization. CA5-1.23 (gamma-1,lambda,titer 3.27x
10000), CB7-1.1(gamma-1,lambda,titer 5.24x100000), CB7-1.5(gamma-
1,lambda,titer 1.31x100000), CB7-1.19 (gamma-1,lambda,titer 1.31x
100000), EA6-2.2 (gamma-1,kappa,titer 5.24x100000),and CB2-2.18 (
mu,kappa,titer 1.31x100000) were further used to preform Western
blotting and neutralization assay. CA5-1.23, CB7-1.1, CB7-1.5,
CB7-1.19, and EE6-2.2 detected a 60KDa protein among the proteins
in Peak 1. However CB2-2.18 detected four proteins with the M.W.
at 60KDa, 51KDa, 50KDa, 47KDa, respectively. In addition, all six
monoclonal antibodies could partially block the P388D1-mediated
tumoricidal activity against MOPC-315 plasmacytoma. The PIF was
further purified using an immunoaffinity column prepared from
CA5-1.23 coupling Affi-Gel 10. Approximately 1045-fold
purification was achieved and the highly purified PIF showed
strong inhibition against the growth of MOPC-315 plasmacytoma at
a dose as low as 0.625ng.
P388D1, macrophage-like cells secreted a plasmacytoma inhibitory