Nikinmaa B, Enns C A, Tonik S E, Sussman H H, Schröder J
Scand J Immunol. 1984 Nov;20(5):441-7. doi: 10.1111/j.1365-3083.1984.tb01023.x.
BALB/c mice were injected with an affinity-chromatography-purified placental transferrin receptor preparation. Spleen cells were fused with NS-1 myeloma cells. Sixteen hybrids producing monoclonal antibodies specific for the transferrin receptor and two hybrids specific for transferrin were identified by radioimmunoassay (RIA). Five hybrids were selected for cloning on the basis of antibody specificity and affinity. None of the antibodies inhibited the binding of transferrin to K562 cells. The binding of antibody ID9 to K562 cells was partially inhibited by transferrin or a polyclonal goat anti-transferrin receptor antiserum. Of the five antibodies, two (IIB6 and IIB2) reacted only with the purified receptor and solubilized cells and not with whole cells. The other three antibodies, when tested with normal human cells and leukaemia and tumour cell lines, showed identical reaction patterns. The antibodies precipitated a glycoprotein from K562 cells with an apparent molecular weight of 94,000, estimated from sodium dodecyl sulphate-polyacrylamide gel electrophoretograms run under reducing conditions, and a molecular weight of 188,000 when run under unreduced conditions. All antibodies have a high affinity with Ka values ranging from 1.44 X 10(9) to 3.56 X 10(10) (l/mol). The antigen precipitated by all five antibodies showed identical peptide maps after partial proteolytic digestion.
将亲和层析纯化的胎盘转铁蛋白受体制剂注射到BALB/c小鼠体内。脾细胞与NS-1骨髓瘤细胞融合。通过放射免疫测定（RIA）鉴定出16种产生针对转铁蛋白受体的单克隆抗体的杂交瘤和2种针对转铁蛋白的杂交瘤。根据抗体特异性和亲和力选择5种杂交瘤进行克隆。没有一种抗体抑制转铁蛋白与K562细胞的结合。转铁蛋白或多克隆山羊抗转铁蛋白受体抗血清可部分抑制抗体ID9与K562细胞的结合。在这5种抗体中，有2种（IIB6和IIB2）仅与纯化的受体和溶解的细胞反应，而不与完整细胞反应。用正常人细胞、白血病和肿瘤细胞系检测时，其他3种抗体表现出相同的反应模式。这些抗体从K562细胞中沉淀出一种糖蛋白，在还原条件下运行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图估计其表观分子量为94,000，在非还原条件下运行时分子量为188,000。所有抗体都具有高亲和力，Ka值范围为1.44×10⁹至3.56×10¹⁰（l/mol）。所有5种抗体沉淀的抗原在部分蛋白酶消化后显示出相同的肽图。
