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用改进的工艺自人心肌组织中提取的高纯度酸性铁蛋白（ＡＩＦ），与经溴化氰（ＢｒＣＮ）活化的ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ偶联，制得ＡＩＦ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ亲和层析柱。用经该柱纯化的兔抗ＡＩＦ抗血清来组建酸性铁蛋白放免分析法（ＡＩＦＲＩＡ）用四参数Ｌｏｇｉｓｔｉｃ数据处理程序处理数据。批内、批间ＣＶ分别为１．６５％和９．７１％，灵敏度为１．８９μｇ／ｌ，回收率为１０２％，可测范围为７．０—３６９．６μｇ／ｌ，ＥＤ５０为２７．５０μｇ／ｌ，与ＡＦＰ、ＣＥＡ、ＬＦ无交叉反应，同常规铁蛋白的交叉反应率为１．７４％。对照组男、女性各３０例，血清ＡＩＦ含量（ｘ±２ｓ）分别为１６．７６±６．９８μｇ／ｌ和１３．３２±６．２５μｇ／１，差异不显著（Ｐ＜０．０５）。１０例肝癌患者血清ＡＩＦ含量为３６．０９±１５．８４μｇ／ｌ，较对照组有非常显著的增高（Ｐ＜０．００１）。上述结果提示，用亲和层析技术纯化ＡＩＦ抗血清是可行的，有利于提高ＲＩＡ的灵敏度、特异性、准确性及临床有效性。
AIF-Sepharose CL-6B affinity chromatography was performed by using an improved process of extracting high purity ferritin (AIF) from human myocardium and coupling with Sepharose CL-6B activated by cyanogen bromide (BrCN). Acid Fractions of Ferritin Assay (AIFRIA) were constructed using rabbit anti-AIF antiserum purified by this column. Data were processed using a four-parameter Logistic data processing program. The intra- and inter-assay CVs were 1.65% and 9.71%, respectively, with a sensitivity of 1.89μg / l and a recovery of 102% with a measurable range of 7.0-369.6μg / l and an ED50 of 27. 50μg / l, no cross-reaction with AFP, CEA, LF, the cross-reactivity with conventional ferritin was 1.74%. There were 30 males and 30 females in the control group. The serum AIF levels (x ± 2s) were 16.76 ± 6.98μg / l and 13.32 ± 6.25μg / l, respectively, with no significant difference (P <0.05). Serum AIF levels in 10 patients with HCC were 36.09 ± 15.84μg / l, which were significantly higher than those in the control group (P <0.001). The above results suggest that the purification of AIF antiserum by affinity chromatography is feasible and will help to improve the sensitivity, specificity, accuracy and clinical efficacy of RIA.