H. Hartmann et al., RAPID QUANTIFICATION OF C3A AND C5A USING A COMBINATION OF CHROMATOGRAPHIC AND IMMUNOASSAY PROCEDURES, Journal of immunological methods, 166(1), 1993, pp. 35-44
Monoclonal antibodies were isolated which reacted specifically with th
e complement cleavage products C3a, C3adR, C5a, and C5adR but not with
the parent molecules C3 or C5. In both cases the mAbs showed a higher
affinity towards the desArg forms. These mAbs were used as capture an
tibodies in immunoassays for C3a/C3adR and C5a/C5adR. The immunoassays
are based on the ABICAP technology which ensures for a rapid measurem
ent. Due to the large binding capacity and the very short diffusion pa
thways in the gel-matrix the binding equilibrium between capture antib
odies and the antigen is reached whilst the sample is flowing through
the column. Therefore this test represents an endpoint assay offering
the possibility of using a single calibration curve for a large number
of measurements. With the C3adR assay concentrations down to 16 ng/ml
C3adR can be detected. The lower detection limit of the C5adR assay i
s 1 ng/ml C5adR. The tests for C3a/C3adR, and C5a/C5adR can be perform
ed in 20 to 25 min and this rapid processing of plasma samples should
permit the application of these parameters for diagnostic purposes and
patient management.