Dübel S, Breitling F, Klewinghaus I, Little M
Recombinant Antibody Group, German Cancer Research Center, Heidelberg.
Cell Biophys. 1992 Aug-Dec;21(1-3):69-79. doi: 10.1007/BF02789479.
A plasmid for optimized protein expression of recombinant Fv antibodies (pOPE) in E. coli was used to express the variable domains of the murine monoclonal antibody HD39 specific for the human B-cell surface antigen CD22. The production of Fv antibodies by pOPE can be regulated over a wide range by varying the IPTG concentration. Antibodies that can discriminate between secreted and nonsecreted Fv antibody fragments were used to show that secretion is the limiting step for the production of functional Fv antibodies. IPTG concentrations above 20 microM increased the total antibody production, but did not yield larger amounts of secreted Fv antibodies. The addition of five histidines to the C terminus facilitates an easy single-step enrichment procedure based on immobilized metal affinity chromatography.
一种用于在大肠杆菌中优化重组Fv抗体（pOPE）蛋白表达的质粒，被用于表达针对人B细胞表面抗原CD22的鼠单克隆抗体HD39的可变区。通过改变异丙基-β-D-硫代半乳糖苷（IPTG）浓度，pOPE产生Fv抗体的过程能够在很宽的范围内受到调控。能够区分分泌型和非分泌型Fv抗体片段的抗体被用于证明分泌是功能性Fv抗体产生的限制步骤。IPTG浓度高于20微摩尔/升时会增加总抗体产量，但不会产生大量分泌型Fv抗体。在C末端添加五个组氨酸有助于基于固定化金属亲和层析的简单单步富集程序。
