NOVEL IMMUNIZATION PROTOCOL AND ELISA SCREENING METHODS USED TO OBTAIN AND CHARACTERIZE MONOCLONAL-ANTIBODIES SPECIFIC FOR HUMAN LIGHT-CHAIN VARIABLE-REGION SUBGROUPS
M. Abe et al., NOVEL IMMUNIZATION PROTOCOL AND ELISA SCREENING METHODS USED TO OBTAIN AND CHARACTERIZE MONOCLONAL-ANTIBODIES SPECIFIC FOR HUMAN LIGHT-CHAIN VARIABLE-REGION SUBGROUPS, Hybridoma, 12(4), 1993, pp. 475-483
We have developed a novel immunization protocol for the production of
a panel of high-affinity murine monoclonal antibodies (MoAbs) that are
specific for each of the major human kappa and lambda light chain var
iable-region (V(L)) subgroups. Mice were injected with heat-precipitat
ed human Bence Jones proteins or V(L)-related fragments emulsified in
monophosphoryl lipid A (MPL) and trehalose dimycolate (TDM) at two- to
four-week intervals over a seven-month period. A unique direct captur
ing enzyme-linked immunosorbent assay (ELISA) employing biotinylated m
onoclonal light chains was designed to select optimally immunized anim
als for hybridoma preparation and to screen culture supernatants for h
igh-affinity anti-V(L) MoAbs. These methods have led to the generation
of MoAbs that by ELISA react specifically with each of the four V(kap
pa) subgroups-V(kappaI), V(kappaII), V(kappaIII), and V(kappaIV) or fi
ve V(lambda) subgroups-V(lambdaI), V(lambdaII/V), V(lambdaIII), V(lamb
daIV), and V(lambdaVI). These reagents have been used successfully to
establish, on the basis of V(L) subgroup, the monoclonal nature of ser
um or urinary immunoglobulins as well as those found in the cytoplasm
or on the cell surface of monoclonal plasma cell or B-lymphocyte popul
ations, respectively. The availability of anti-V(L) subgroup-specific
MoAbs will facilitate the immunodiagnosis and study of patients with m
ultiple myeloma, AL amyloidosis, and related B-cell proliferative diso
rders.