人源性抗大疱性类天疱疮抗原BP180-NC16A单链抗体的构建及表达  

Construction and Expression of Human Anti-BP180-NC16A Single-chain Antibody

作　　者：孙娜娜[1] 王欲晓[2] 周丽君[2] 王刚[1] 

机构地区：[1]第四军医大学西京医院全军皮肤病研究所,陕西西安710032 [2]海军总医院中心实验室,北京100037

出　　处：《中国皮肤性病学杂志》2009年第2期71-74,共4页The Chinese Journal of Dermatovenereology

摘　　要：目的用基因工程技术制备人源性抗大疱性类天疱疮抗原BP180单链抗体(scFv),并对其抗原结合特异性等进行鉴定。方法利用基因重组技术对从噬菌体抗体库中克隆的人源性抗大疱性类天疱疮抗原BP180-NC16A抗体Fab片段进行改造获得scFv,进行DNA测序,同时用ELISA法鉴定其抗原结合活性和特异性,免疫荧光技术检测scFv与人皮肤组织BP180抗原的结合情况。结果成功构建了抗BP180-NC16A scFv,并获得可溶性表达,DNA测序结果表明,单链抗体的Vλ和VH基因序列完全正确,ELISA法证实可溶性表达的scFv抗体具有良好的抗原结合活性和特异性。免疫荧光检测该抗体在人皮肤表皮、真皮交界处基底膜带呈现线状结合条带。结论成功构建并表达了人源性抗大疱性类天疱疮抗原BP180-NC16A的单链抗体,为进一步研究该抗体的生物学作用奠定了基础。Objective To construct human single -chain Fv antibody （scFv） against bullous pemphigoid antigen BP180- NC16A and identify it＇s binding activity and specificity. Methods By genetic engineering technology, human anti-BP180-NC16A scFv was constructed from Fab antibody fragment selected from phage antibody library. The binding activity and specificity of the expressed scFv products were identified by ELISA and indirect immunofluorescence （IIF）. Results DNA sequence analysis showed that there was no any mutation in Vλ and VH genes of the constructed scFv. The soluble anti-BP180-NC16A scFv was suc cessfully expressed and found to have a good antigenic specificity as well as excellent binding activity with target antigen by ELISA and showed linear deposition along the dermal-epidermal junction （DEJ） by IIF. Conclusion Human anti-BP180-NC16A scFv was successfully expressed. It would be useful in furthering the studies on the pathogenesis and treatment of BP.

关 键 词：大疱性类天疱疮 BP180 单链抗体 

分 类 号：R758.66[医药卫生—皮肤病学与性病学]