Foti M, Granucci F, Ricciardi-Castagnoli P, Spreafico A, Ackermann M, Suter M
CNR Cellular and Molecular Pharmacology Center, Milano, Italy.
J Immunol Methods. 1998 Apr 15;213(2):201-12. doi: 10.1016/s0022-1759(98)00029-5.
Rabbit monoclonal antibodies (RmAb) are not routinely obtained by eukaryotic cell fusion techniques. Therefore, we have applied phage display technology to produce a recombinant rabbit Fab molecule directed against the KLH model antigen. The Fab fragments selected from the rabbit phage display library were subcloned in an expression vector to permit the production of a fusion protein comprising a dimer of bacterial alkaline phosphatase (phoA). This fusion protein was directly produced into the periplasmic space of Escherichia coli. We show that a crude extract containing these conjugates can be used in a direct enzyme immunoassay, as exemplified in the case of the KLH antigen.
兔单克隆抗体(RmAb)通常不是通过真核细胞融合技术获得的。因此,我们应用噬菌体展示技术来生产针对钥孔血蓝蛋白(KLH)模型抗原的重组兔Fab分子。从兔噬菌体展示文库中筛选出的Fab片段被亚克隆到表达载体中,以产生包含细菌碱性磷酸酶(phoA)二聚体的融合蛋白。这种融合蛋白直接在大肠杆菌的周质空间中产生。我们证明,含有这些偶联物的粗提物可用于直接酶免疫测定,如在KLH抗原的例子中所示。