用于测定培养上清液和人血清中鼠单克隆抗体的定量酶联免疫吸附测定。

Quantitative ELISA for mouse monoclonal antibody determination in culture supernatants and in human serum.

作者信息

Vilaseca J C, Pupo M, Bernal M, Matamoros L, Gordillo S, Rodríguez H, Caballero Y, Otero A

机构信息

Institute of Tropical Medicine Pedro Kouri, Ciudad de la Habana, Cuba.

出版信息

Hybridoma. 1997 Dec;16(6):557-62. doi: 10.1089/hyb.1997.16.557.

DOI:10.1089/hyb.1997.16.557

PMID:9455710

Abstract

An ELISA has been set up for quantifying mouse monoclonal antibodies in culture supernatant. The assay includes rabbit anti-mouse IgG antibodies chromatographycally purified. This preparation was used as coating and as conjugated antibodies in the ELISA. The assay can detect IgG1 with sensitivity of 0.2 ng/mL, IgG2a (0.85 ng/mL), IgG2b (0.13 ng/mL), and IgG3 (3.19 ng/mL) in culture supernatants. The effective working range was from subnanogram per mL quantities to 30 ng/mL by using a computer statistical program. Variation coefficient of ELISA was below 7%. Correlation estimates with a similar ELISA using commercial reagents were performed for each mouse antibody subclass. The assay was able to detect the four mouse monoclonal antibody subclasses in pure human serum as compared with the same ELISA using commercial antibodies. A 24-h pharmacokinetic profile of 1 patient treated with an IgG2a monoclonal antibody is presented.

摘要

已建立一种酶联免疫吸附测定法（ELISA），用于定量培养上清液中的小鼠单克隆抗体。该测定法包括经色谱纯化的兔抗小鼠IgG抗体。此制剂用作ELISA中的包被抗体和偶联抗体。该测定法能检测培养上清液中的IgG1，灵敏度为0.2 ng/mL，IgG2a（0.85 ng/mL），IgG2b（0.13 ng/mL）和IgG3（3.19 ng/mL）。通过使用计算机统计程序，有效工作范围为每毫升亚纳克量至30 ng/mL。ELISA的变异系数低于7%。针对每种小鼠抗体重链亚类，与使用商业试剂的类似ELISA进行了相关性评估。与使用商业抗体的相同ELISA相比，该测定法能够在纯人血清中检测到四种小鼠单克隆抗体重链亚类。展示了1例接受IgG2a单克隆抗体治疗患者的24小时药代动力学曲线。