Troianovskiĭ S M, Krutovskikh V A, Bannikov G A
Biull Eksp Biol Med. 1986 Jun;101(6):733-6.
BALB/c mice were immunized with intermediate filaments (IF) from the rat colon mucosa, and their splenocytes were fused with myeloma cells to obtain hybridomas. Specific antibody production was assessed by indirect immunofluorescence on cultured rat hepatoma 27 containing prekeratins. The clones that stained IF in hepatoma and not in fibroblasts were judged positive. Clones E3 and E6 were shown to produce monoclonal antibodies against prekeratin with molecular mass of 40 kD (PK40), while clones E2 and E7 produced antibodies against prekeratin with molecular mass of 55 kD (PK55). This was established by immunoblotting with 125I-protein A in cell lysates from the colon, bladder, and hepatoma 27. Only PK55 was revealed in liver and salivary gland lysates. The above proteins were not detected in esophagus, fibroblast and skeletal muscle cell lysates. The monoclonal antibodies make it possible to study individual prekeratin expression in embryogenesis, differentiation and neoplastic transformation of simple epithelium.
用大鼠结肠黏膜的中间丝（IF）免疫BALB/c小鼠，然后将其脾细胞与骨髓瘤细胞融合以获得杂交瘤。通过对含有前角蛋白的培养大鼠肝癌27细胞进行间接免疫荧光来评估特异性抗体的产生。在肝癌细胞中能染色IF而在成纤维细胞中不能染色的克隆被判定为阳性。已证明克隆E3和E6产生针对分子量为40kD的前角蛋白（PK40）的单克隆抗体，而克隆E2和E7产生针对分子量为55kD的前角蛋白（PK55）的抗体。这是通过用125I-蛋白A对来自结肠、膀胱和肝癌27细胞的裂解物进行免疫印迹确定的。在肝脏和唾液腺裂解物中仅检测到PK55。在食管、成纤维细胞和骨骼肌细胞裂解物中未检测到上述蛋白质。这些单克隆抗体使得研究简单上皮细胞在胚胎发生、分化和肿瘤转化过程中单个前角蛋白的表达成为可能。
