한국산 겨우살이 렉틴(KML-C)에 대한 단일크론항체의 생산과 특징

Production of Monoclonal Antibodies Specific to Korean Mistletoe Lectin(KML-C) and Their Characterization

We have reported that water-extracted Korean mistletoe(KM-110) had various biological activities such as antitumor and immunomodulatory activity and the lectin fraction(KML-C) of the extract was one of the major factors related to its biological functions. In this paper, we produced murine monoclonal antibody(mAb) against KML-C. The mAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a lectin from European mistletoe. One group mABs(9H7-D10 and 3C2-1H4) strongly reacted with KML-C, but not ML-I. In contrast, another group mAbs(8B11-2C5, 8E12-3E9 and 5E10-F1) reacted with both KML-C and ML-1. The subisotypes of these mAbs were shown to be IgG1(9H7-1D10, 3C2-1H4 and 8B11-2C5) or IgM(8E12-3E9 and 5E10-F1). To develop an assay system for determination of the amonunt of KML-C, we established the sandwich ELISA(enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase(HRP) labelled mAbs.In various combinations of the mAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C,showing the detection limit ranging from 7-5,000ng/ml. Especially, reproducibility(C.V.) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8B11-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.