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Generation and characterization of recombinant scFv antibodies detecting Eimeria acervulina surface antigen

Cited 15 time in Web of Science Cited 16 time in Scopus
Authors
Kim, Jin-Kyoo; Min, Wongi; Lillehoj, Hyun Soon; Kim, Sungwon; Sohn, Eun Jung; Song, Ki Duk; Han, Jae Yong
Issue Date
2001
Publisher
Mary Ann Liebert
Citation
Hybridoma, vol.20 no.3, pp. 175-181
Abstract
In our previous attempt to generate monoclonal antibodies (MAbs) against coccidia parasites that more accurately
reflect the natural avian humoral immune response, we produced two chicken B-cell hybridomas,
5D11 and 2-1. While both cell lines secreted antibodies reactive with sporozoites of Eimeria acervulina, they
were produced in yields too low to conduct meaningful in vivo studies. To circumvent this problem, we produced
four single chain variable fragment (scFv) antibodies from the VH and VL genes of hybridomas 5D11
and 2-1. The concentration of these recombinant antibodies expressed in E. coli and purified to homogeneity
was 5–6 mg/L. Three of the antibodies exhibited antigen binding specificity to Eimeria surface antigens equivalent
to that of the native MAbs. Nucleotide sequence analysis of the VL genes from hybridomas 5D11 and 2-
1 and genomic DNA revealed vestiges of gene conversion with Vl pseudogenes. These recombinant scFv antibodies
will prove useful for further characterization of natural Eimeria surface antigens as potential vaccine
candidates.
ISSN
1554-0014
Language
English
URI
https://hdl.handle.net/10371/99038
DOI
https://doi.org/10.1089/027245701750293501
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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