Kodama S, Kishi J, Obata K, Iwata K, Hayakawa T
Fuji Chemical Industries, Ltd., Takaoka, Japan.
Coll Relat Res. 1987 Oct;7(5):341-50. doi: 10.1016/s0174-173x(87)80027-4.
Hybridoma antibodies against bovine collagenase inhibitor were produced by fusion of myeloma cells NS-1 (P3-NS1-1) with spleen cells from mice hyperimmunized with collagenase inhibitor purified from the explant medium of bovine dental pulps. Hybridomas positive by an enzyme-linked immunosorbent assay (ELISA) for bovine collagenase inhibitor were cloned by the dilution method. Seventeen hybridomas producing antibodies were isolated, four of which also recognized purified human collagenase inhibitor in the ELISA. Using a monoclonal antibody-Sepharose affinity column, we easily purified both bovine and human collagenase inhibitors to homogeneity. They showed the same mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, corresponding to a molecular mass of 32,000 daltons.
通过将骨髓瘤细胞NS-1（P3-NS1-1）与用从牛牙髓外植体培养基中纯化的胶原酶抑制剂高度免疫的小鼠脾细胞融合，产生了抗牛胶原酶抑制剂的杂交瘤抗体。通过稀释法克隆了经酶联免疫吸附测定（ELISA）对牛胶原酶抑制剂呈阳性的杂交瘤。分离出17个产生抗体的杂交瘤，其中4个在ELISA中也识别纯化的人胶原酶抑制剂。使用单克隆抗体-琼脂糖亲和柱，我们轻松地将牛和人胶原酶抑制剂纯化至同质。它们在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上显示出相同的迁移率，对应于32,000道尔顿的分子量。
