Biomedical Engineering Reference
In-Depth Information
FIGURE 12.5
Molecular imaging of internalized SHG2210:mAb 35 complex in human muscle
cells by confocal microscopy SHG2210 fusion protein and mAb 35 were directly labeled with Alexa
fluor 546 and 488 dyes, respectively. Fusion protein and antibody were preincubated for 15min to
form complexes and then added to the human muscle cell line DB40 for 2 h on ice. Cells were then
washed and switched to 37
C to initiate uptake of the complex. Uptake of complex was allowed to
occur for 45 min, at which point cells were fixed and processed for immunofluorescence using anti-
Lamp1 monoclonal antibody. Clockwise from top: mAb 35 (green); SHG2210 (red); Lamp-1 (blue);
merged image.
SHG2210 was absent [19]. Data from a typical experiment
are presented in Figure 12.6.
compartments after internalization; however, neither
SHG2210 nor SHG2210/antibody complex is present in
Rab-11 positive recycling endosomes. Thus, SHG2210 and
SHG2210/anti-AchR complex is degraded with negligible
recycling observed. Significantly, SHG2210 is shown to have
a protective effect on antigenic modulation of the AChR
induced by serum from selected patients withMG, suggesting
that a fusion protein approach may be an effective therapeutic
for treating MG.
12.3.6 Summary of In Vitro Studies of SHG2210
In vitro studies were performed to assess the efficacy of
SHG2210. Key assays and technology platforms include
cell-based binding and uptake assays, molecular imaging,
and confocal microscopy studies and antigenic modulation
assays. In vitro studies show (1) SHG2210 binds to anti-AChR
antibodies, (2) SHG2210:antibody complexes are specifically
targeted to the transferrin receptor on cells, and (3) fusion
protein:antibody complex is taken into cells and traffics
to the lysosome. The SHG2210 and SHG2210/anti-AchR
antibody complex is present in Lamp-1 positive lysosomal
12.4 APPLICATIONS AND INDICATIONS
SHG2210 is designed as a specific therapeutic for MG.
Indeed, it is even specific to a certain subset of MG patients:
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