一种用于同时免疫组织化学筛选96个杂交瘤克隆以产生组织特异性抗体的新方法，筛选出了一种针对昆虫心侧体的单克隆抗体。

A new alternative for simultaneous immunohistochemical screening of 96 hybridoma clones for tissue-specific antibody productions selects a monoclonal antibody to insect corpus cardiacum.

作者信息

Verhaert P, De Loof A, Huybrechts R, Delang I, Theunis W, Clottens F, Schoofs L, Swinnen K, Vandesande F

出版信息

J Neurosci Methods. 1986 Sep;17(4):261-8. doi: 10.1016/0165-0270(86)90126-3.

Abstract

In the current search for the elucidation of the true structure of hitherto unidentified 'new' insect neuropeptides we designed a novel screening method to facilitate the primary detection of neurone-specific antibody secreting mouse-mouse hybridoma clones obtained after immunization with neuronal tissue homogenates. The present procedure is principally adapted from a conventional immunohistological test and enables one to rapidly screen 96 (and even more) clones at one time for potential secretion of specific antibodies to different tissue compounds, without the necessity of having a purified antigen. It has proved to be sensitive, rapid, practical and reproducible. As such it promises to be very useful to discriminate amongst the wide range of antibodies to various kinds of materials produced by hybridomas by detecting monoclonal antibodies directed against factors contained in well-defined tissues in which one is interested. This paper also reports the successful application of this method to a primary screening of clones producing murine monoclonal antibodies to substances of insect corpora cardiaca (CC), after immunization with crude antigen preparations.

摘要

在当前致力于阐明迄今尚未鉴定的“新”昆虫神经肽真实结构的研究中，我们设计了一种新型筛选方法，以促进对用神经组织匀浆免疫后获得的神经元特异性抗体分泌型小鼠 - 小鼠杂交瘤克隆进行初步检测。本方法主要改编自传统免疫组织学检测方法，能够一次性快速筛选96个（甚至更多）克隆，以检测针对不同组织成分的特异性抗体的潜在分泌情况，而无需纯化抗原。事实证明，该方法灵敏、快速、实用且可重复。因此，通过检测针对感兴趣的特定组织中所含因子的单克隆抗体，有望在区分杂交瘤产生的针对各种物质的广泛抗体方面非常有用。本文还报道了该方法在用粗抗原制剂免疫后，成功应用于对产生抗昆虫心侧体（CC）物质的鼠单克隆抗体的克隆进行初步筛选。