抗人TNF-α单链抗体rScFvW13B1原核表达、纯化及功能鉴定

Prokaryotic Expression and Purification of Anti-hTNF-αScFv

应用PET32-c原核表达载体快速、高效地表达及纯化单链抗体。将单链抗体W13B1基因克隆到PET32-c原核表达载体中,通过酶切及测序鉴定正确后,进行原核诱导表达并纯化,并检测纯化得到的单链抗体的功能。DNA琼脂糖凝胶电泳表明,单链抗体基因克隆成功;SDS-PAGE结果表明,单链抗体得到成功表达和纯化;ELISA和WesternBlot结果表明,单链抗体W13B1能够特异性结合人TNF-α。说明可成功地表达及纯化抗人TNF-α单链抗体rScFv-W13B1。

To express and purify the ScFv antibody rapidly and effectively by applying PET32-c vector, the gene of rScFv W13B1 was cloned into PET32-c vector. After the detection of enzyme digestion and sequencing, the prokaryotic expression and purification were processed and the purified ScFv was detected by ELISA and Western blot. The gene of ScFv was cloned rightly. The analysis of SDS-PAGE showed the ScFv was expressed and purified successfully. The result of ELISA and Western blot indicated the rScFvW13B1 could bind hTNF-α specifically. It is conclused that the prokaryotic expression and purification of rScFv-W13B1 were established successfully.