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Quantitation of angiogenesis factor in bovine retina and tumour extracts by means of radioimmunoassay.

机译:通过放射免疫测定定量牛视网膜和肿瘤提取物中的血管生成因子。

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摘要

Using an antiserum raised against tumour angiogenesis factor (TAF) we have developed a radioimmunoassay for retina and tumour angiogenesis factor(s). This antiserum was previously shown to bind to both human and animal tumour extracts and to inhibit the angiogenesis induced by TAF in vivo. TAF from rat Walker tumour was used for iodination by the chloramine-T method. An excess of 125I-labelled TAF was incubated with TAF antibody in the absence (maximum binding) and presence (inhibition of maximum binding) of unlabelled tissue extract. A double antibody technique was used to separate free and bound TAF. Unlabelled human Wilms tumour TAF was used as a standard. The extent of inhibition of 125I-TAF-anti-TAF binding provided a measure of TAF in tissue extracts examined. Extracts of normal bovine retina, cornea, lung, aorta, lymph nodes, iris, vitreous humour, and human tumours and normal human pituitary and liver were assayed. Only bovine retina and human tumours were found to contain angiogenesis factor. These findings, together with our earlier results, suggest that angiogenesis factor from both bovine retina and human tumours induce angiogenesis in vivo and possess common antigenic determinants. The presence of angiogenesis factor in healthy retina and its relationship to neovascularisation in clinical conditions is discussed.
机译:使用针对肿瘤血管生成因子(TAF)产生的抗血清,我们开发了一种针对视网膜和肿瘤血管生成因子的放射免疫测定法。先前已证明该抗血清与人和动物肿瘤提取物结合并在体内抑制TAF诱导的血管生成。通过氯胺-T法将来自大鼠Walker肿瘤的TAF用于碘化。在未标记的组织提取物不存在(最大结合)和存在(最大结合抑制)的情况下,将过量的125 I标记的TAF与TAF抗体一起孵育。使用双重抗体技术来分离游离和结合的TAF。以未标记的人类Wilms肿瘤TAF为标准。 125 I-TAF-抗TAF结合的抑制程度提供了所检测组织提取物中TAF的量度。分析了正常牛视网膜,角膜,肺,主动脉,淋巴结,虹膜,玻璃体液和人肿瘤以及正常人垂体和肝脏的提取物。仅发现牛视网膜和人肿瘤含有血管生成因子。这些发现以及我们先前的结果表明,来自牛视网膜和人肿瘤的血管生成因子均在体内诱导血管生成,并具有常见的抗原决定簇。讨论了健康视网膜中血管生成因子的存在及其与临床条件下新血管形成的关系。

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