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应用常规方法制备了４株能稳定分泌抗人重组γ－干扰素单克隆抗体（ＭｃＡｂ）的小鼠杂交瘤细胞系：３Ｄ３、３Ｄ１２、３Ｆ９和３Ｂ８．经免疫琼脂双扩散法鉴定，其分泌的ＭｃＡｂ的亚类分别为：ＩｇＧ２ｂ、ＩｇＧ３、ＩｇＧ１和１ｇＧ１．相对亲和常数前两株ＭｃＡｂ为６．２５×１０－１１、后两株ＭｃＡｂ为６．２５×１０－１３．问接ＥＬＩＳＡ法测培养上清的效价为２×１０２～１．２８×１０５．由小鼠腹水提取的４株ＭｃＡｂ与天然γ－干扰素均产生交叉反应，对重组的人γ－干扰素均具有中和活性．其中３Ｆ９和３Ｄ３培养上清对重组人γ－干扰素亦有明显的中和活性。用３Ｆ９ＭｃＡｂ与溴化氢活化的ＳｅＰｈａｒｏｓｅ４Ｂ偶联制备了亲和层析柱，用其纯化粗提的γ－干扰素纯度达到９５％以上，比活性达１．９×１０７ＩＵ／ｍｇ蛋白．
Four mouse hybridoma cell lines stably secreting anti-human recombinant γ-interferon monoclonal antibody (McAb) were prepared by conventional methods: 3D3, 3D12, 3F9 and 3B8. Identified by immunogold agar double diffusion method, the subclasses of McAb secreted were IgG2b, IgG3, IgG1 and IgG1 respectively. Relative affinity constant before the two McAb 6.25 × 10-11, the latter two McAb 6.25 × 10-13. The titer of the culture supernatant was 2 × 102 ~ 1.28 × 105. The 4 McAbs extracted from mouse ascites produced cross-reaction with natural γ-interferon and had neutralizing activity on recombinant human γ-interferon. The 3F9 and 3D3 culture supernatants also showed significant neutralizing activity on recombinant human interferon-γ. Affinity chromatography was carried out by coupling 3F9McAb with hydrogen bromide-activated SePharose4B. Purified crude interferon-γ reached more than 95% purity with a specific activity of 1.9 × 107IU / mg protein.