Pedersen L O, Andreasen R B
Institute of Experimental Immunologi, Copenhagen, Denmark.
Hybridoma. 1989 Feb;8(1):97-105. doi: 10.1089/hyb.1989.8.97.
An IgG human-human monoclonal hybridoma antibody, AML-19, reactive with human myeloid cells of non-malignant and malignant origin has been produced by fusion of blood mononuclear cells from a patient with acute myeloid leukemia (AML) and the human B-lymphoma cell line RH-L4. The monoclonal antibody (MAb) AML-19 was purified from hybridoma supernatant by primarily anion-exchange chromatography, in order to separate the AML-19 MAb from contaminating immunoglobulin (Ig), e.g. bovine Ig and MAb derived from the parental fusion partner, and followed by immunoaffinity chromatography. This purification method gave the highest yield and purity of the AML-19 MAb. The isoelectric point (pI) of the MAb was estimated to be 5. Inhibition assays indicate an apparent dissociation constant (Kd) corresponding to 4 x 10(-9) M and an affinity constant (Ka) to 2.5 x 10(8)M-1 to K562 erythroleukemia cells. Scatchard plot demonstrated a linear slope as a manifestation of monoclonality and a low number of AML-19 specific epitopes, estimated to 1500 per cell.
一种与非恶性和恶性来源的人类髓细胞发生反应的IgG型人-人单克隆杂交瘤抗体AML-19，是通过将一名急性髓细胞白血病（AML）患者的血液单核细胞与人类B淋巴瘤细胞系RH-L4融合产生的。单克隆抗体（MAb）AML-19首先通过阴离子交换色谱法从杂交瘤上清液中纯化，以便将AML-19单克隆抗体与污染的免疫球蛋白（Ig），如牛Ig和源自亲本融合伙伴的单克隆抗体分离，然后进行免疫亲和色谱法。这种纯化方法得到了最高产量和纯度的AML-19单克隆抗体。该单克隆抗体的等电点（pI）估计为5。抑制试验表明，其与K562红白血病细胞的表观解离常数（Kd）对应于4×10⁻⁹ M，亲和常数（Ka）为2.5×10⁸ M⁻¹。Scatchard图显示出线性斜率，这是单克隆性的表现，且AML-19特异性表位数量较少，估计每个细胞有1500个。
