De Leij L, Poppema S, The T H
J Immunol Methods. 1983 Aug 12;62(1):69-72. doi: 10.1016/0022-1759(83)90111-4.
A new freezing technique is described which permits time-consuming protocols as a first screening of newly formed hybridomas. In this procedure complete 96 well clustertrays with growing hybridomas are cryopreserved after programmed freezing. This procedure has been successfully applied to a number of fusion protocols for which the objective was to obtain monoclonal antibodies against tissue specific antigens. To this end hybridoma supernatants were screened by an immunoperoxidase technique on frozen sections. Freezing of the hybridoma containing clustertrays permitted extensive screening and partial characterization of the previously collected supernatants. After subsequent thawing of appropriate wells, the hybridoma clones proved to be viable and usually no loss of antibody production was observed.
本文描述了一种新的冷冻技术，该技术允许采用耗时的方案作为对新形成的杂交瘤的初步筛选。在此过程中，将装有生长中的杂交瘤的完整96孔细胞培养板在程序冷冻后进行冷冻保存。该程序已成功应用于许多融合方案，其目的是获得针对组织特异性抗原的单克隆抗体。为此，通过免疫过氧化物酶技术在冷冻切片上筛选杂交瘤上清液。含有杂交瘤的细胞培养板的冷冻允许对先前收集的上清液进行广泛筛选和部分表征。在随后对合适的孔进行解冻后，杂交瘤克隆被证明是有活力的，并且通常未观察到抗体产生的损失。
