Jk. Kim et al., Generation and characterization of recombinant ScFv antibodies detecting Eimeria acervulina surface antigens, HYBRIDOMA, 20(3), 2001, pp. 175-181
In our previous attempt to generate monoclonal antibodies (MAbs) against co
ccidia parasites that more accurately reflect the natural avian humoral imm
une response, we produced two chicken B-cell hybridomas, 5D11 and 2-1. Whil
e both cell Lines secreted antibodies reactive with sporozoites of Eimeria
acervulina, they were produced in yields too low to conduct meaningful in v
ivo studies, To circumvent this problem, we produced four single chain vari
able fragment (scFv) antibodies from the VH and VL genes of hybridomas 5D11
and 2-1. The concentration of these recombinant antibodies expressed in E.
coil and purified to homogeneity was 5-6 mg/L. Three of the antibodies exh
ibited antigen binding specificity to Eimeria surface antigens equivalent t
o that of the native MAbs, Nucleotide sequence analysis of the VL genes fro
m hybridomas 5D11 and 2-1 and genomic DNA revealed vestiges of gene convers
ion with Vh pseudogenes, These recombinant scFv antibodies will prove usefu
l for further characterization of natural Eimeria surface antigens as poten
tial vaccine candidates.