Reactivities of monoclonal antibodies with E. bieneusi spores
MAb | Result with:b | |||||
---|---|---|---|---|---|---|
Frozen MAba | Spore fixation method | Deglycosylation (periodate) | Deglycosylation (enzymatic)c | |||
Methanol | Paraformaldehyde | Acetone | ||||
1D4 | ++ | + | ++++ | ++++ | ++++ | ++++ |
3E9 | ++++ | ++ | ++ | ++++ | ++++ | ++++ |
2B2 | ++++ | + | ++ | ++++ | ++++ | ++++ |
2G4 | ++++ | + | − | ++++ | ++++ | ++++ |
2E9 | ++++ | + | ++ | ++++ | ++++ | ++++ |
4B2 | ++++ | + | − | ++++ | ++++ | ++++ |
4C9 | ++++ | ++ | ++ | ++++ | ++++ | ++++ |
4C3 | ++++ | + | ++ | ++++ | ++++ | ++++ |
↵a Reactivities of the MAbs frozen at −70°C were tested after every month for 3 months. Reactivity intensities remained the same during that period.
↵b +, faint fluorescence intensity; ++, weak fluorescence intensity; +++, strong fluorescence intensity; ++++, very strong fluorescent intensity.
↵c Enzymatic deglycosylation removed N-linked, simple O-linked, and complex O-linked carbohydrates but did not alter reactivity of any MAb even after 5 days of deglycosylation.