Development of single chain variable fragment (scFv) antibodies against surface proteins of ‘Ca. Liberibacter asiaticus’

Serological detection of ‘Candidatus Liberibacter asiaticus’ in citrus, and identification by GeLC-MS/MS of a chaperone protein responding to cellular pathogens

“…Single chain variable fragment antibodies scFv 748 (OmpA) and scFv 932 (FlgI) were selected for testing in the DTBIA. These scFv had been used successfully when expressed from phagemid vector pKM19 to detect purified protein antigens and in dot blot assays against crude extracts from infected sweet orange 21 . In the present work the scFv were expressed from the plasmid vector pKM16 in E. coli BL21 and purified by taking advantage of the 6X His tag provided by the vector.…”

“…A library of single chain antibodies (scFv) with 2 × 10 7 unique members was created using the phagemid vector pKM19 and Escherichia coli DH5αF’ for screening and selection, and plasmid pKM16 for propagation of the scFv 21 25 . This scFv library was screened to select scFv that bound to segments of proteins expressed on the surface of C aLas, including the major outer membrane protein, OmpA (scFv748; YP_003065185) and a membrane component of the flagellar apparatus, FlgI (scFv932; YP_003064789).…”

“…This library of single chain antibodies (scFv) was made using phage display technology, and includes unique mouse antibodies that recognize antigens present in psyllids infected by C aLas that were used to immunize mice as the first step in the creation of the library. scFv are available that recognize antigens expressed on the surface of C aLas cells, including the major outer membrane protein, OmpA and a protein component of the basal flagellar apparatus, FlgI 21 .…”

Production of monoclonal antibody against Ompa protein of Candidatus Liberibacter asiaticus causing citrus greening disease

“…Yuan và đtg (2016) đã sử dụng kháng thể đơn chuỗi scFv kháng một số kháng nguyên của vi khuẩn CLas, tuy nhiên chỉ có kháng thể đơn chuỗi kháng kháng nguyên Ompa cho kết quả tốt nhất trong việc phát hiện mẫu bệnh vàng lá gân xanh bằng phản ứng ELISA. Tuy vậy, sự sai khác của giá trị OD450 trong phản ứng ELISA giữa mẫu bị nhiễm bệnh vàng lá gân xanh và mẫu khỏe là chưa lớn (Yuan et al, 2016). Như vậy, cần có các nghiên cứu khác để tối ưu hiệu quả của kháng thể đơn chuỗi trong việc phát hiện bệnh vàng lá gân xanh bằng kỹ thuật ELISA.…”

A Review of the Citrus Greening Research and Development Efforts Supported by the Citrus Research and Development Foundation

“…The 5-year $9 million NuPsyllid project formulated by CRDF and funded by USDA NIFA concluded in 2017. The very ambitious overall goal of the project, to develop a modified psyllid that would not transmit CLas and could be deployed in the field to displace the natural ACP population, was not achieved but the project did contribute significant advances to the fundamental knowledge of ACP-CLas biology (Soderlund et al, 2014;Cicero et al, 2015;Ding et al, 2015Ding et al, , 2016Ding et al, , 2017Yuan et al, 2015Yuan et al, , 2016Brown et al, 2016;Stover et al, 2016;Liu et al, 2017).…”