人体体液中人表皮生长因子——hEGF水平的放射免疫测定

Radioimmunoassay of human epidermal growth factor--hEGF levels in human body fluids.

作者信息

Hayashi T, Sakamoto S

机构信息

Central Research Laboratories, Wakunaga Pharmaceutical Co., Ltd., Hiroshima, Japan.

出版信息

J Pharmacobiodyn. 1988 Mar;11(3):146-51. doi: 10.1248/bpb1978.11.146.

DOI:10.1248/bpb1978.11.146

PMID:3261789

Abstract

By immunizing rabbits with purified human epidermal growth factor (hEGF), which was synthesized by Escherichia coli constructed by a genetic engineering technique, high titer antiserum for hEGF was obtained. The antiserum has an excellent specificity to hEGFs (hEGF, hEGF [1-51], hEGF [1-47] and [Leu21] hEGF) and has almost no cross-reactivity with other biological peptides, such as mouse EGF and human platelet derived growth factor. Using radioiodinated hEGF, a radioimmunoassay (RIA) for hEGF was established by a double antibody-polyethylene glycol method. As little as 10 pg/tube of hEGF was successfully determined by the present RIA methods. The intra- and inter-assay coefficients of variation for hEGF in human plasma or serum were 2.4-4.9% and 3.7-6.2%, respectively. The hEGF levels in human body fluids determined by the present method were as follows: urine (11-100 ng/ml), serum (1.0-1.8 ng/ml), plasma (0.2-0.8 ng/ml), sperm (17-47 ng/ml), saliva (0.9-3.0 ng/ml) and tears (9.5-27 ng/ml).

摘要

用通过基因工程技术构建的大肠杆菌合成的纯化人表皮生长因子（hEGF）免疫家兔，获得了高滴度的hEGF抗血清。该抗血清对hEGF（hEGF、hEGF [1-51]、hEGF [1-47] 和 [Leu21] hEGF）具有优异的特异性，与其他生物肽，如小鼠EGF和人血小板衍生生长因子几乎没有交叉反应。使用放射性碘标记的hEGF，通过双抗体-聚乙二醇法建立了hEGF的放射免疫分析（RIA）。本RIA方法成功测定的hEGF低至10 pg/管。人血浆或血清中hEGF的批内和批间变异系数分别为2.4-4.9%和3.7-6.2%。用本方法测定的人体体液中的hEGF水平如下：尿液（11-100 ng/ml）、血清（1.0-1.8 ng/ml）、血浆（0.2-0.8 ng/ml）、精子（17-47 ng/ml）、唾液（0.9-3.0 ng/ml）和泪液（9.5-27 ng/ml）。