流行性出血热免疫球蛋白的研制

本文首次报告采用纯化灭活流行性出血热（ＥＨＦ）Ⅰ型疫苗免疫健康献血员，采集ＥＨＦ抗体高滴度的血浆，用低温乙醇法及盐析法分离纯化三批ＥＨＦ免疫球蛋白。结果表明：（１）采用０，１，３，４（月）或０，１，４，５（月）免疫程序，疫苗剂量１～２ｍｌ（０．１５～０．３０ｍｇ蛋白），受免献血员血清平均抗体滴度可达１∶４０６（ＥＬＩＳＡ）或１∶１１２（ＲＰＨＩ）。（２）通过生化检定，三批制品的电泳纯度为９７．０５％，９６．８４％，９９．２６％；ＩｇＧ单体和二聚体含量为８９．５５％，９１．３０％和９８．２１％。（３）用空斑抑制中和试验及免疫印染试验证明所纯化的免疫球蛋白具有抗ＥＨＦ病毒特异性。（４）三批ＥＨＦ免疫球蛋白的效价测定，结果为ＥＬＩＳＡ滴度≥１∶５１２，ＲＰＨＩ滴度≥１∶１０２４，ＰＲＮＴ（中和抗体）滴度１∶４０。按１６％蛋白计，ＥＨＦ免疫球蛋白的效价可达原料血浆的１０倍以上。（５）无菌、安全、毒性及热原质试验检定结果，全部通过《中国生物制品规程》要求。

Study on the Preparation of Human Plasma lmmunoglobulin of EHF

A study on the preparation of human plasma immunoglobulin of epidemic hemorrhagic fever (EHF)was reported. The normal donors were immunized with the purified and inactivated EHF vaccine (type Ⅰ, strain LR1), then the immune plasma of highly specific antibody was collected from these donors. Three lots of EHF immunoglobulins were prepared successfully from above immunoplasma by both methods of cold alcohol and(NH4)2SO4 salting out. The results indicated:(1)the anti-EHF titers of serum reached 1 :406 (ELISA) and 1:112 (RPHI) respectively; (2)the electrophoresis purity of the preparations was up to 97% to 99%, and the monomer and dimer contents of IgG were 89.55%,91.30% and 98.21% by sephadex gel chrormatography; (3)the products were specific to EHF antigen by PRNT and Western-blot test:(4)the titers of the EHF immunoglobulin were more than 1 :512(ELISA), 1:1024 (RPHI) and 1:40 (PRNT) respectively, and the potency of final products was about tenfold higher than that of original immuno-plasma; (5)all of the sterlity. safety, abnormal toxicity and pyrogenicity tests for the preparations met the Chinese Requirements of Biological Products (1990) .