As a member of the tumor necrosis factor (TNF) superfamily, the
B-cell activating factor (BAFF) plays a crucial role in B-cell survival
and differentiation. Overexpression of this protein has been closely
linked to autoimmune disorders and some B-cell malignancies. Using
monoclonal antibodies (mAbs) against the BAFF soluble domain appears to
be a complementary treatment for some of these diseases. This study
aimed to produce and develop a specific Nanobody (Nb), a variable
camelid antibody domain, against the soluble domain of BAFF protein.
After camel immunization with recombinant protein and preparing cDNA
from total RNAs separated from camel lymphocytes, an Nb library was
developed. Individual colonies capable of binding selectively to rBAFF
were obtained by periplasmic-ELISA, sequenced, and expressed in a
bacterial expression system. The specificity and affinity of selected Nb
were determined and its target identification and functionality were
evaluated using flow cytometry.