TABLE 3.
bvMSP142 | % Final puritya | Endotoxin level (U/mg)b | Mass (kDa) determined by:
|
N-terminal sequenced | Glycosylation pattern | Monoclonal antibody recognitione | |
---|---|---|---|---|---|---|---|
Gel filtration chromatography | Electron spray mass spectroscopyc | ||||||
Secreted | 97.4 | 0.78 | 81.5f | 43,320.0; 44,404.0f | NH2 ADPAVTPSVIDNIL | High mannose | 5.2, 2D634, 13E353, 4H919, 12.1, 1E1, 12.8, 2F10 |
Intracellular | ND | ND | 100.0 | 41,320; 42,276; 43,660 | NH2 ADPAVTPSVIDNIL | High mannose | Same as for secreted bvMSP142 |
Determined by scanning densitometry. ND, not determined.
Determined by Limulus amebocyte lysate assay.
Major mass peaks are listed.
ADP is a vector-derived cloning sequence; underlining indicates the start of the MSP142 sequence. The predicted sequence is NH2 ADPAVT.
Monoclonal antibodies recognizing conformational epitopes in MSP119 with different specificities were used in Western blot analyses: 5.2 (32), 2D634, 13E353, 4H919 (7), 12.1 (25), 1E1, 12.8, and 2F10 (16). Antibodies 5.2 and 12.8 recognize the first EGF domain; 4H919 recognizes the second EGF domain; and 2D634, 13E353, 12.1, and 12.8 recognize both EGF domains.
Predicted: 41,924.4.