二胺氧化酶抗血清制备及夹心法ELISA测定

Production of Antiserum Against Diamine Oxidaseand Its Application to Sandwith ELISA

将二胺氧化酶(DAO)与福氏佐剂乳化后,采用淋巴结免疫法制得兔抗DAO抗血清,双扩法测其效价为1∶32。再经盐析法及DEAE-32离子交换法层析得到抗DAO-IgG,并用辣根过氧化物酶以过碘酸钠法标记,建立了DAO的ELISA检测方法,并对方法的灵敏度,精确度和特异性作了研究。测定了113例献血员血清中DAO含量((?)±s),其中53例男性为0.27±0.24μg/ml,60例女性为0.45±0.33μg/ml。结果显示男女性别间有显著差异(P<0.01)。

A rabbit antiserum angainst diamine oxidase (DAO) was prpared by the injections of the emulsified antigen, which was that DAO was emulsified with Freund's adjuvant into lymph mode. The antiserum titer was 1:32 by double diffusion. Immunoglobulin G fractions were obtained by salting out and DEAE - 32 ion - exchange chro-matography, and were lablled with HRP by the oxidation method of sodium periodate. Using the antibody, a sand-with ELISA for DAO was developed to determine the DAO content in serum. 113 valunteers were observed, the rusults revealed that the DAO content of 53 male is 0.27 - 0.24ug/ml( X±S), that of 60 female is 0.45 ± 0.33μg/ ml. The result showed that the difference between male and female is significant (P<0.01). inthe meantime, the sensitivity and precision of the method were analysed.