Noll F, Handschack W, Eichmann E, Gaestel M, Schneider F, Kuhl R, Löster K
Central Institute of Molecular Biology, Academy of Sciences of GDR, Berlin-Buch.
Biomed Biochim Acta. 1989;48(1):165-76.
Four monoclonal antibodies against HuIFN-alpha-1 and -alpha-2 were produced by the lymphocyte hybridoma technique. The screening for monoclonal anti-IFN antibodies was performed by two different procedures permitting the detection of neutralizing and non-neutralizing antibodies. The first antibody screening in the supernatants of hybridoma clones was carried out by an immunodot ELISA using a novel chromogen, 2-bromo-1-naphthol, for the immunoperoxidase reaction. The second screening was performed with a solid-phase bioassay. The monoclonal antibodies IIG11 and IVH2 react specifically with HuIFN-alpha-1 independently of its origin, natural or recombinant. The mAb IIG11 recognizes also an epitope on a 27 kDa interferon, probably HuIFN-alpha-8. The mAb IIG7 binds to HuIFN-alpha-2 only, whereas the mAb IIB2 binds to HuIFN-alpha-2 and also to the 27 kDa interferon and to a 25 kDa interferon, not yet further characterized. These antibodies can be used successfully for immunopurification as well as for quantification of the corresponding interferons.
采用淋巴细胞杂交瘤技术制备了4种抗人干扰素α-1和α-2的单克隆抗体。通过两种不同的方法筛选单克隆抗干扰素抗体,这两种方法可检测中和抗体和非中和抗体。杂交瘤克隆上清液中的首次抗体筛选是通过免疫斑点ELISA进行的,该方法使用新型显色剂2-溴-1-萘酚进行免疫过氧化物酶反应。第二次筛选采用固相生物测定法。单克隆抗体IIG11和IVH2能特异性与人干扰素α-1反应,与其来源(天然的或重组的)无关。单克隆抗体IIG11还识别一种27 kDa干扰素上的表位,可能是人干扰素α-8。单克隆抗体IIG7仅与人干扰素α-2结合,而单克隆抗体IIB2与人干扰素α-2结合,还与27 kDa干扰素和一种25 kDa干扰素结合,该25 kDa干扰素尚未进一步鉴定。这些抗体可成功用于相应干扰素的免疫纯化和定量分析。