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目的 获得鼠抗人精浆蛋白单抗（ｍＡｂ）κ链基因，并将其转染到ＨｅＬａ细胞进行表达。方法 从分泌抗人精浆蛋白ｍＡｂ的杂交瘤细胞系Ｅ４Ｂ７中提取总ＲＮＡ，用ＲＴ－ＰＣＲ法获取κ链ｃＤＮＡ。将其克隆入真核表达载体ｐｃＤＮＡ３，用脂质体转染法转染ＨｅＬａ细胞，并用免疫荧光染色法检测κ链的表达。结果从分泌抗人精浆蛋白的鼠ｍＡｂ杂交瘤细胞系Ｅ４Ｂ７中克隆了κ链基因。序列测定表明，Ｖκ属于鼠免疫球蛋白ⅩⅢ家族。以含κ链基因的真核表达载体ｐｃＤＮＡ３－Ｅ４Ｂ７κ转染ＨｅＬａ细胞后，在ＨｅＬａ细胞中检测到了κ链的表达。结论 获得了序列正确的κ链基因，并在ＨｅＬａ细胞中成功地表达，为进一步构建和表达Ｆａｂ段打下了良好基础。
Objective To obtain the murine anti-human seminal plasma monoclonal antibody (mAb) κ chain gene and transfect it into HeLa cells for expression. Methods Total RNA was extracted from the hybridoma cell line E4B7 secreting anti-human seminal plasma mAb and the κ-chain cDNA was obtained by RT-PCR. The recombinant plasmid was cloned into the eukaryotic expression vector pcDNA3 and transfected into HeLa cells by lipofectamine 2000. The expression of κ chain was detected by immunofluorescence staining. Results The κ chain gene was cloned from murine mAb hybridoma cell line E4B7 secreting anti-human seminoprotein. Sequence analysis showed that VK belongs to murine immunoglobulin XIII family. After the HeLa cells were transfected with the eukaryotic expression vector pcDNA3-E4B7κ containing κ-chain gene, the expression of κ-chain was detected in HeLa cells. Conclusion The correct kappa chain gene was obtained and successfully expressed in HeLa cells, which laid a good foundation for further construction and expression of Fab fragment.