用双特异性单克隆抗体对牛乳铁蛋白进行竞争性酶联免疫吸附测定

Competitive ELISA of bovine lactoferrin with bispecific monoclonal antibodies.

作者信息

Shinmoto H, Kobori M, Tsushida T, Shinohara K

机构信息

National Food Research Instituted, Ministry of Agriculture, Forestry, and Fisheries, Ibaraki, Japan.

出版信息

Biosci Biotechnol Biochem. 1997 Jun;61(6):1044-6. doi: 10.1271/bbb.61.1044.

DOI:10.1271/bbb.61.1044

PMID:9214770

Abstract

A mouse hybrid-hybridoma, HH1-4-3, secreting IgG1 class bispecific antibodies to bovine lactoferrin (bLF) and horseradish peroxidase (HRPO), was established previously. The competitive enzyme-linked immunosorbent assay (ELISA) of bLF using the HH1-4-3 culture supernatant was not sensitive enough to measure bLF concentration in biological fluids. To improve the sensitivity of the competitive ELISA, we fractionated the bispecific antibodies by antigen affinity column chromatography. A column immobilized with bLF adsorbed 60% of the antibodies of the HH1-4-3 supernatant, and the amount of antibodies adsorbed on a column immobilized with HRPO was less than 5%. The competitive ELISA of bLF using the affinity purified bispecific antibodies through an HRPO-immobilized column chromatography showed a good standard curve at bLF concentrations of 10 ng/ml to 100 micrograms/ml.

摘要

先前已建立了一种小鼠杂交 - 杂交瘤HH1 - 4 - 3，它能分泌针对牛乳铁蛋白（bLF）和辣根过氧化物酶（HRPO）的IgG1类双特异性抗体。使用HH1 - 4 - 3培养上清液对bLF进行竞争性酶联免疫吸附测定（ELISA），其灵敏度不足以测量生物体液中的bLF浓度。为提高竞争性ELISA的灵敏度，我们通过抗原亲和柱色谱法对双特异性抗体进行了分级分离。用bLF固定的柱子吸附了HH1 - 4 - 3上清液中60%的抗体，而用HRPO固定的柱子上吸附的抗体量不到5%。使用通过HRPO固定柱色谱法亲和纯化的双特异性抗体对bLF进行竞争性ELISA，在bLF浓度为10 ng/ml至100微克/毫升时显示出良好的标准曲线。