Vollmers H P, Wozniak E, Stepien-Bötsch E, Zimmermann U, Müller-Hermelink H K
Institut für Pathologie, Universität Würzburg, Germany.
Hum Antibodies Hybridomas. 1996;7(1):37-41.
We describe in this paper the rapid and cheap purification of human immunoglobulin M from hybridoma supernatant of mass culture. The method consists of two steps: 1. concentration of supernatant by ultrafiltration and 2. dialysis against distilled water, pH 6.4. To produce the supernatant, the hybridomas are grown in RPMI media with fetal calf serum (10% FCS) in 250 ml flasks under normal tissue culture conditions. More than 14 mg IgM can be nearly selectively purified within 6 h from 5 l of antibody containing hybridoma supernatant. Most of the IgM molecules stay in a penta- and monomeric form and are positive in physiological and immunohistochemical studies.
我们在本文中描述了从大规模培养的杂交瘤上清液中快速且廉价地纯化人免疫球蛋白M的方法。该方法包括两个步骤：1. 通过超滤浓缩上清液；2. 用pH 6.4的蒸馏水进行透析。为了获得上清液，杂交瘤在正常组织培养条件下，于含有胎牛血清（10% FCS）的RPMI培养基中，在250毫升烧瓶中培养。在6小时内，可从5升含抗体的杂交瘤上清液中近乎选择性地纯化出超过14毫克的IgM。大多数IgM分子保持五聚体和单体形式，并且在生理学和免疫组织化学研究中呈阳性。
