Rodríguez-Martínez Luis Mario, Marquez-Ipiña Alan Roberto, López-Pacheco Felipe, Pérez-Chavarría Roberto, González-Vázquez Juan Carlos, González-González Everardo, Trujillo-de Santiago Grissel, Ponce-Ponce de León César Alejandro, Zhang Yu Shrike, Dokmeci Mehmet Remzi, Khademhosseini Ali, Alvarez Mario Moisés
Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Monterrey, Nuevo León, México.
Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Monterrey, Nuevo León, México; Biomaterials Innovation Research Center, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America; Harvard-Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
PLoS One. 2015 Oct 21;10(10):e0135859. doi: 10.1371/journal.pone.0135859. eCollection 2015.
Current Ebola virus (EBOV) detection methods are costly and impractical for epidemic scenarios. Different immune-based assays have been reported for the detection and quantification of Ebola virus (EBOV) proteins. In particular, several monoclonal antibodies (mAbs) have been described that bind the capsid glycoprotein (GP) of EBOV GP. However, the currently available platforms for the design and production of full-length mAbs are cumbersome and costly. The use of antibody fragments, rather than full-length antibodies, might represent a cost-effective alternative for the development of diagnostic and possibly even therapeutic alternatives for EBOV.
METHODS/PRINCIPAL FINDINGS: We report the design and expression of three recombinant anti-GP mAb fragments in Escherichia coli cultures. These fragments contained the heavy and light variable portions of the three well-studied anti-GP full-length mAbs 13C6, 13F6, and KZ52, and are consequently named scFv-13C6, scFv-13F6, and Fab-KZ52, respectively. All three fragments exhibited specific anti-GP binding activity in ELISA experiments comparable to that of full-length anti-GP antibodies (i.e., the same order of magnitude) and they are easily and economically produced in bacterial cultures.
CONCLUSION/SIGNIFICANCE: Antibody fragments might represent a useful, effective, and low cost alternative to full-length antibodies in Ebola related capture and diagnostics applications.
目前的埃博拉病毒(EBOV)检测方法成本高昂,在疫情场景中不实用。已报道了不同的基于免疫的检测方法用于埃博拉病毒(EBOV)蛋白的检测和定量。特别是,已经描述了几种结合EBOV糖蛋白(GP)衣壳的单克隆抗体(mAb)。然而,目前用于设计和生产全长单克隆抗体的平台繁琐且成本高昂。使用抗体片段而非全长抗体,可能是开发EBOV诊断甚至治疗替代方案的一种经济有效的选择。
方法/主要发现:我们报道了三种重组抗GP单克隆抗体片段在大肠杆菌培养物中的设计和表达。这些片段包含三种经过充分研究的抗GP全长单克隆抗体13C6、13F6和KZ52的重链和轻链可变部分,因此分别命名为scFv-13C6、scFv-13F6和Fab-KZ52。在ELISA实验中,所有这三种片段均表现出与全长抗GP抗体相当的特异性抗GP结合活性(即相同数量级),并且它们可以在细菌培养物中轻松且经济地生产。
结论/意义:在埃博拉相关的捕获和诊断应用中,抗体片段可能是全长抗体的一种有用、有效且低成本的替代方案。