The rapid generation of high-quality target antibodies is essential for research employing immunoassays. The use of recombinant antibody technology that relies on genetic engineering is one such means to produce high-quality antibodies. Obtaining the gene sequence information of immunoglobulin is a prerequisite for the preparation of genetically engineered antibodies. At present, many researchers have shared their amino acid sequence data for various high-performance antibodies and their related properties. In this study, we obtained the protein sequence of a variable region of a 17 β-estradiol (E₂) antibody from the Protein Data Bank (PDB) and subsequently constructed heavy (H) and light (L) chain expression vectors through codon optimization. The transient expression, purification, and performance identification of the immunoglobulin G (IgG), antigen-binding fragment (Fab), and single-chain variable fragment (scFv) antibodies were carried out, respectively. The effects of the different expression vectors on the expression yield of the IgG antibody were further compared. Among them, the expression yield based on the pTT5 vector was the highest, reaching 27 mg/L. Based on the expressed IgG and Fab antibodies, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) standard curve of E₂ was constructed, and the half-maximal inhibitory concentrations (IC₅₀) for these two antibodies were determined to be 0.129 ng/mL and 0.188 ng/mL, respectively. In addition, an immunochromatographic assay (ICA) based on the IgG antibody was constructed with an IC₅₀ of 3.7 ng/mL. Therefore, in featuring the advantages of simplicity, high efficiency, rapid obtainment, and high titer yield, we propose the system for the rapid generation of high-quality recombinant antibodies by reusing the published antibody information and show that it has good implementation prospects in improving upon existing immunoassay techniques.

快速生成高质量的目标抗体对于采用免疫测定的研究至关重要。使用依赖于基因工程的重组抗体技术是生产高质量抗体的一种手段。获得免疫球蛋白的基因序列信息是制备基因工程抗体的先决条件。目前，许多研究人员已经分享了他们针对各种高性能抗体及其相关特性的氨基酸序列数据。在这项研究中，我们获得了 17 β-雌二醇（E ₂) 来自蛋白质数据库 (PDB) 的抗体，随后通过密码子优化构建了重链 (H) 和轻链 (L) 表达载体。分别进行了免疫球蛋白G（IgG）、抗原结合片段（Fab）和单链可变片段（scFv）抗体的瞬时表达、纯化和性能鉴定。进一步比较了不同表达载体对IgG抗体表达量的影响。其中，基于pTT5载体的表达量最高，达到27 mg/L。_{以表达的IgG和Fab抗体为基础，构建了E 2}的间接竞争酶联免疫吸附试验（ic-ELISA）标准曲线，半数抑制浓度（IC ₅₀) 这两种抗体的浓度分别为 0.129 ng/mL 和 0.188 ng/mL。此外，构建了基于 IgG 抗体的免疫层析分析 (ICA)，其 IC ₅₀为 3.7 ng/mL。因此，我们以简单、高效、快速获取、高效价等优点为基础，提出了利用已发表的抗体信息快速生成高质量重组抗体的系统，并表明其在改进方面具有良好的实施前景。基于现有的免疫测定技术。